Journal of the International Academy of Periodontology
Transcription
Journal of the International Academy of Periodontology
Journal of the International Academy of Periodontology The official journal of the International Academy of Periodontology Volume 13Number 1 January 2011 Published by Stephen Hancocks Ltd JIAP JIAP Jan Oct 11 10 OFC.indd OFC.indd 11 22/12/2010 23/09/2010 10:21:43 13:01:19 INFORMATION FOR AUTHORS Manuscripts Manuscripts for publication and all correspondence should be sent to Dr. Mark R. Patters, Editor, Journal of the International Academy of Periodontology, University of Tennessee College of Dentistry, Office of Academic Affairs, 875 Union Avenue, Memphis, TN USA, 38163, e-mail: JIAP@uthsc.edu. Printed submissions sent to the above address must be prepared as described below and accompanied by a standard floppy disk containing an electronic copy of the manuscript. The floppy disk should be labelled with the manuscript title, author(s) and specific version of the word processing program used. In lieu of printed manuscripts, electronic submissions done entirely in Microsoft Word (PC or Mac) will be accepted at the above e-mail address. Effective 1 January 2006, the corresponding author of a manuscript submitted for publication in the Journal of the International Academy of Periodontology must be a member of the International Academy of Periodontology or, in lieu of membership, pay a submission fee of US$100. An application for membership can be found at http://www.perioiap.org/join_iap.htm. Those authors choosing to pay the submission fee should contact the Editor at JIAP@uthsc.edu before submitting the manuscript.. All submissions must be written in English and will be subject to peer and editorial review. Articles for publication will be considered under the following headings: original research, clinical case reports and review articles relevant to all aspects of periodontology and implantology. Articles must be original and may not have been submitted or accepted for publication elsewhere, with the exception of presentation at a scientific meeting and publication as an abstract. A signed statement to this effect should be included with the submission of the manuscript. Research that involves studies on humans must conform to the Declaration of Helsinki and the authors must indicate that appropriate informed consent was obtained. Research reports State the problem and objectives clearly, describe the methods and materials in detail, report the results clearly using the minimum number of figures and tables; and, bearing in mind previously published work, discuss the results, the conclusions, and the clinical implications. Clinical case reports Discuss a clinical challenge; describe the treatment method and discuss the results in light of previously published methods of treatment of individual patients. Literature reviews Record the sequence of development of a particular aspect of periodontology in detail, as briefly and succinctly as possible. The review should cover the topic completely and be thoroughly referenced. At least one contributing author of a review must have personal experience with relevant research. Letters to the Editor Letters may address relevant matters of concern to the membership of the International Academy of Periodontology or offer constructive criticism of articles published by JIAP. Letters must be concise and signed. If the letter comments on a published article, it should contain appropriate references. The letter will be referred to the author(s) of the original work so that they will have an opportunity to respond. Editorials Editorials may be solicited from authorities to provide a unique perspective on published articles, or to comment on other items of interest to the membership. Copyright statement A copyright transfer statement will accompany the galley proofs of accepted, typeset manuscripts. The form must be signed by at least one of the authors and returned with the corrected proofs. Manuscript preparation Manuscripts must be submitted in quadruplicate (one original and three copies) and should be typewritten, double-spaced on one side only of A4 or 8.5 x 11 inch paper, with at least 25 millimetre (1 inch) margins on all four sides. Articles generally should not exceed 10–12 pages (excluding references, tables, figure legends and figures) and should be limited to no more than six authors. Additional contributing authors will be listed as an addendum to the manuscript. Abbreviations should be placed in parenthesis after the first complete use of the term(s) to be abbreviated. Use generic names for drugs and for dental materials. Give trade names and manufacturers’ names and addresses in parentheses. 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Please provide three to six key words (Dental Descriptors, Index to Dental Literature and/or Index Medicus) to be used for indexing purposes. Text The body of the manuscript should contain an Introduction, a detailed review of the Materials and Methods, a description in logical sequence of Results, and a Discussion section with Conclusions. Acknowledgements and Conflict of Interest Include acknowledgement of those individuals who contributed to the publication, source of financial support, and any financial relationships of any of the authors which may pose a perceived conflict of interest. References In the text the author’s (authors’) name(s) and date of publication should be used as either: “in a similar study (Anderson and Morgan, 1992)”, or “Conversely, Blinkhorn (1994) found that.” If there are more than two authors, the first author and year are cited in the text; for example, “(Spencer et al., 1995)”. Citation of authors of more than one paper in a single year are shown as 1995a; 1995b; etc. Multiple references in the text should appear in chronological order separated by semicolons. Authors of unreferenced work should appear in the text only. The list of references at the end of the text should be double-spaced and arranged alphabetically by name of first author. All authors should be listed unless there are more than six, when only the first three should be given followed by et al. This should be followed by the title of the article, the full name of the journal (in italics); the year of publication; the volume number (in bold); and the first and last page numbers in full. Please follow the punctuation used in the examples below. Papers submitted with an incoreectly formatted reference list will be held without review until a corrected reference list is provided. Examples: Reference to an article: Shiloah, J. and Patters, M.R. Repopulation of Periodontal Pockets by Microbial Pathogens in the Absence of Supportive Therapy. Journal of Periodontology 1996; 67:130–139. Reference to a book: Schuster, G.S. Oral Microbiology and Infectious Diseases, 3rd ed. Philadelphia: B. C. Decker, 1990; 516–522. Reference to a chapter in a book: Chesney, J., Patters, M.R. and Budreau-Patters, A. Oral Infections. In Long, S., Prober, C. and Pickering, L. (Eds): Principles and Practice of Pediatric Infectious Disease. New York. John Wiley and Sons, 1996. Reference to a dissertation or thesis: James, A. On the Immune Response to GTR Membranes in Periodontics. PhD, Liverpool, UK. 1994; 15–25. Reference to a report: Committee on Mercury Hazards in Dentistry. Code of Practice for Dental Mercury Hygiene. London: Department of Health and Social Security, 1979; Publication No. DHSS 79-F372. Reference to an abstract: Patters, M.R., Shiloah, J., Dean, J.W., Bland, P. and Toledo, G. The Consequence of Infection of Treated Periodontal Pockets by Microbial Pathogens. Journal of Dental Research 1997; 76 (special issue), 111 (Abst). Tables Each table must be submitted on a separate sheet of paper, double spaced throughout, including column heads, footnotes, and data. They should be numbered with arabic numerals according to their order of mention in the text. Tables should be self-explanatory and supplement, not duplicate, the text. All footnotes should immediately follow the table, and all abbreviations should be defined in the footnote. Illustrations Illustrations should be numbered with arabic numerals in order of their mention in the text. They may be submitted in colour or black and white, but colour will be used at the Editor’s discretion where it enhances the text. In general, the Editor will require that clinical photographs and stained histologic specimens be submitted and published in colour. 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Authors must disclose to the Editor any financial interest they may have in products mentioned in their article. The Journal will endeavour to provide a decision to the authors within 16 weeks of arrival of the paper at the editorial office, provided that the corresponding author has a functioning e-mail address. Reliance on overseas mail may delay the decision from reaching the author within the above time frame. Manuscripts submitted for review that do not follow these instructions may be delayed until corrected or returned unreviewed. Volume 13 Number 1 January 2011 ISSN 1466–2094 EDITORIAL BOARD Mark R Patters Editor Memphis, TN, USA Andrea B Patters Associate Editor Sultan Al Mubarak Riyadh, Saudi Arabia P Mark Bartold Adelaide, SA, Australia Michael Bral New York, NY, USA Nadine Brodala Chapel Hill, NC, USA Cai-Fang Cao Beijing, People’s Republic of China Chong-Pyoung Chung Seoul, Korea Daniel Etienne Paris, France Journal of the International Academy of Periodontology The Effect of Different Interdental Cleaning Devices on Gingival Bleeding Nanning A. M. Rosema, Nienke L. Hennequin-Hoenderdos, Claire E. Berchier, Dagmar E. Slot, Deborah M. Lyle and Godefridus A. van der Weijden 2 Additive or Synergistic Antimicrobial Effects of Amoxicillin and Metronidazole on Whole Plaque Samples: A Preliminary Report Clemens Walter, Eva M. Kulik, Roland Weiger, Nicola U. Zitzmann and Tuomas Waltimo 11 Clinical and Microbiological Comparison of Three Non-surgical Protocols for the Initial Treatment of Chronic Periodontitis Carlos Serrano, Nidia Torres, Angela Bejarano, Marcela Caviedes and María Eugenia Castellanos 17 13th International Biennial Congress - Registration Form 27 Erhan Firatli Istanbul, Turkey Kohji Hasegawa Tokyo, Japan Vincent J Iacono Stony Brook, NY, USA Isao Ishikawa Tokyo, Japan Georges Krygier Paris, France Yoji Murayama Okayama, Japan Hamdy Nassar Cairo, Egypt Angela R C Pack Dunedin, New Zealand David Paquette Chapel Hill, NC, USA Stephen Polins Boston, MA, USA Rok Schara Ljubljana, Slovenia Lior Shapira Jerusalem, Israel Uros Skaleric Ljubljana, Slovenia Aubrey Soskolne Jerusalem, Israel Thomas E Van Dyke Boston, MA, USA JIAP Jan 11 Contents pp1.indd 97 The Journal of the International Academy of Periodontology is the official journal of the International Academy of Periodontology and is published quarterly (January, April, July and October) by Stephen Hancocks Ltd in association with Dennis Barber Ltd. Manuscripts, prepared in accordance with the Information for Authors should be submitted to the Editor, Dr. Mark R. Patters, University of Tennessee, Department of Periodontology, 875 Union Avenue, Memphis, TN 38163, USA. Electronic submissions will be accepted and should be sent to jiap@utmem.edu. Instructions to Authors are available at: http://www.perioiap.org/publications.htm#JOURNAL All enquiries concerning advertising, subscriptions, inspection copies and back issues should be addressed to Mrs. Kelly Ogilvie McLean, Goldman School of Graduate Dentistry, Boston University, 100 E. Newton Street, Boston, MA, USA 02118. Tel: +1 617 638-4758; Fax: +1 617 638-4799. Email: bujiap@bu.edu. Whilst every effort is made by the publishers and Editorial Board to see that no inaccurate or misleading opinion or statement appears in this Journal they wish to make clear that the opinions expressed in the articles, correspondence, advertisements etc., herein are the responsibility of the contributor or advertiser concerned. Accordingly, the publishers and the Editorial Board and their respective employees, offices and agents accept no liability whatsoever for the consequences of any such inaccurate or misleading opinion or statement. © 2011 International Academy of Periodontology. All rights reserved. No part of this publication may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, or otherwise, without permission of the Academy. Produced in Great Britain by Dennis Barber Limited, Lowestoft, Suffolk 23/12/2010 10:18:46 Journal of the International Academy of Periodontology 2011 13/1: 2–10 The Effect of Different Interdental Cleaning Devices on Gingival Bleeding Nanning A. M. Rosema1, Nienke L. Hennequin-Hoenderdos1, Claire E. Berchier1, Dagmar E. Slot1, Deborah M. Lyle2 and Godefridus A. van der Weijden1 Department of Periodontology, Academic Centre for Dentistry Amsterdam ACTA, University of Amsterdam and VU University Amsterdam,The Netherlands, and 2Water Pik Inc, Fort Collins, CO, USA 1 Abstract Objective: To compare the effectiveness of an oral irrigator (OI) with a prototype jet tip or a standard jet tip to floss as adjunct to daily toothbrushing on gingival bleeding. Methods: In this single masked, 3-group parallel, 4-week home use experiment, 108 subjects were randomly assigned to one of three groups: 1) OI with a prototype jet tip; 2) OI with a standard jet tip; 3) waxed dental floss. All groups used their assigned product once a day as adjunct to twice daily toothbrushing for two minutes with a standard ADA reference toothbrush. Professional instructions were given by a dental hygienist in OI use or floss use according to written instructions. All subjects also received a toothbrush instruction leaflet (Bass technique). Subjects were assessed for both bleeding and plaque at baseline and after two weeks and four weeks and were instructed to brush their teeth approximately 2 to 3 hours prior to their assessment. Results: With respect to mean bleeding scores the ANCOVA analysis with baseline as covariate and week 4 as dependent variable showed a significant difference between groups in favor of both the oral irrigator groups. For plaque, however, no significant difference among groups was observed. Conclusion: When combined with manual toothbrushing the daily use of an oral irrigator, either with prototype or standard jet tip, is significantly more effective in reducing gingival bleeding scores than is the use of dental floss, as determined within the limits of this 4-week study design. Key words: Floss, dental water jet, oral irrigator, water flosser, gingivitis, bleeding, plaque, toothbrush Introduction Biofilms are 3-dimensional arrangements of bacteria that are loosely or more firmly adherent to teeth and tissue (Costerton et al., 1994). Biofilms consist of microcolonies of bacteria embedded in slimy matrices and are self-sufficient, dynamic communities that can survive in hostile environments (Marsh and Bradshaw, 1995) The regular removal of dental plaque biofilm, which contains the bacteria responsible for caries formation and for the etiology of gingivitis and periodontitis, is the wellaccepted conditio sine qua non of dental health (Gorur et al., 2009). Mechanical removal is considered the most effective method to control the growth of the oral biofilm. The most common device used for mechanical plaque control Correspondence to: N.A.M. Rosema. Department of Periodontology, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Gustav Mahlerlaan 3004 1081 LA Amsterdam, The Netherlands. E-mail: M.Rosema@acta.nl is either a manual or power toothbrush. As toothbrush efficacy is limited to the surfaces of the teeth it can access (facial, lingual, and occlusal), another device is needed to clean the interdental area and the proximal surfaces of the teeth and surrounding gingivae. Other factors that affect the efficacy of mechanical plaque biofilm removal include brushing frequency, brushing time, toothbrush design, and brushing technique (Jepsen et al., 1998; Van der Weijden et al., 1993). For most people, however, total plaque biofilm removal is not a realistic goal. It is difficult for patients to effectively remove or disrupt the biofilm from all surfaces of the teeth on a daily basis (Douglass et al., 1993; Brown et al., 1993). On average, people reduce their plaque scores by approximately 50% by brushing (Jepsen et al., 1998). Therefore, compliance with instructions is a major consideration when recommending any self-care device. To be truly patient-centered, practitioners must shift to recommending available tools that, besides having demonstrated efficacy in reducing inflammation based on scientific evidence, are also preferred by patients (Slot et al., 2008). © International Academy of Periodontology JIAP 10-005 Rosema.indd 2 04/01/2011 11:37:43 The Effect of Different Interdental Cleaning Devices on Gingival Bleeding A dental water jet or water flosser or oral irrigator (OI) is an electric device that delivers a pulsating fluid via controlled pressure which is aimed at the removal of interdental and subgingival plaque biofilm on tooth surfaces to reduce inflammation as a supplement to toothbrushing (Lobene, 1969; Drisko et al., 1987; Cobb et al., 1988; Flemmig et al., 1990; Chaves et al., 1994; Flemmig et al.,1995; Barnes et al., 2005; Gorur et al., 2009). The OI was introduced to the dental profession in 1962 and has been studied extensively for the past decades. Clinical studies demonstrate that an OI is safe and can significantly reduce bleeding and gingivitis in a variety of cohorts (Lobene et al., 1969; Flemmig et al., 1990; Brownstein et al., 1990; Burch et al., 1994; Newman et al., 1994; Flemmig et al., 1995; Felo et al., 1997; Barnes et al., 2005; Sharma et al., 2008). However, erythrosinebased plaque indices have yielded equivocal data. Some studies have shown a reduction in plaque indices with the use of the OI compared to toothbrushing alone (Burch et al., 1994; Felo et al., 1997; Cutler et al., 2000; Al Mubarak et al., 2002; Sharma et al., 2008), while other studies showed no significant differences (Ciancio et al, 1989; Brownstein et al., 1990; Walsh et al., 1992; Chaves et al., 1994; Fine et al., 1994). The OI is likely to provide a particular benefit in terms of gingival health to a large part of the general public that does not clean the interproximal spaces on a regular basis (Research, Science and Therapy Committee, 2005). In 2001 the American Academy of Periodontology stated, “Among individuals who do not perform excellent oral hygiene, supragingival irrigation with or without medicaments is capable of reducing gingival inflammation beyond that normally achieved by toothbrushing alone. This effect is likely due to the flushing out of subgingival bacteria (Research, Science and Therapy Committee, 2001).” In a 2005 position paper, the American Academy of Periodontology stated that “supragingival lavage can assist individuals with gingivitis or poor oral hygiene. The greatest benefit is seen in patients who perform inadequate interproximal cleansing. Patients report that the OI facilitates the removal of food debris in posterior areas, especially in cases of fixed bridges or orthodontic appliances, when the proper use of interdental cleaning devices is difficult” (Research, Science and Therapy Committee, 2005). However, anecdotal discussions and commentary continue concerning the appropriate use and efficacy of this instrument. OI devices can be used with water but also with antimicrobial agents (Flemmig et al., 1990, Brownstein et al., 1990, Jolkovsky et al., 1990; Newman et al., 1994, Fine et al., 1994; Chaves et al., 1994; Flemmig et al., 1995; Felo et al., 1997). The objective of the present study was to test the adjunctive effect to toothbrushing of an OI with either a prototype jet tip or a standard tip in the potential to JIAP 10-005 Rosema.indd 3 3 improve gingival health over a 4-week period. This prototype tip, which is configured with filaments, may help the user guide the tip along the gingival margin and the interproximal area. Both OI tips were compared to the use of dental floss. These treatments were combined with the use of a regular flat trimmed manual toothbrush together with a standard dentifrice. Materials and methods Study population One hundred seventy-two subjects (non-dental students) from different universities and colleges in and around Amsterdam responded to an e-mail advertisement and reported for a screening appointment. The volunteers were informed about the study, first in a recruitment letter and secondly at the screening. Participation was not limited by race or gender. Subjects received a written explanation of the background of the study, its objectives and their involvement. Before screening for their suitability they were all requested to give their written informed consent. Subjects were required to fulfill the following criteria: ≥18 years of age, a minimum of five evaluable teeth in each quadrant (with no partial dentures, orthodontic banding or wires); moderate gingivitis (50% bleeding on marginal probing, Galgut et al., 1998), an absence of oral lesions and/or periodontal pockets > 5 mm and/or generalized recession, and the absence of pregnancy and systemic diseases such as AIDS, cirrhosis, diabetes, any adverse medical history or long-term medication, or any physical condition that limits manual dexterity. All subjects received oral and written information about the products and purpose of the study. One hundred eight subjects met the inclusion criteria and were enrolled into the study, which was conducted in accordance to the ethical principles that have their origin in the Declaration of Helsinki and was consistent with Good Clinical Practice guidelines. Medical Ethics Committee approval was obtained prior to the start of the study (MEC 09/198 #09.17.1322). All assessments took place at the Department of Periodontology at ACTA, Amsterdam, The Netherlands in September and October, 2009. Study products Three different interdental products were tested in this study, one product per group, with 36 subjects enrolled in each group. All subjects received a standard toothbrush (Oral-B Indicator 35, Procter & Gamble, Cincinnati, OH, USA, Figure 1) and standard fluoride dentifrice (Everclean, HEMA, Amsterdam, The Netherlands). In addition, subjects were randomized (see below for details) into one of three groups for assignment of an interdental cleaning device: 04/01/2011 11:37:43 4 Journal of the International Academy of Periodontology (2011) 13/1 Group 1 (OIP): OI (DWJ-Waterpik® Ultra Water Flosser, Fort Collins, CO, USA) with a prototype jet tip (Figure 2, test group). Group 2 (OIS): OI (DWJ-Waterpik® Ultra Water Flosser, Fort Collins, CO, USA) with a standard jet tip (Figure 3, benchmark control group, Husseini et al., 2008). Group 3 (DF): standard waxed floss (Johnson & Johnson, New Brunswick, NJ, USA) (Figure 4, control group). Clinical assessment Clinical parameters were assessed at baseline (S1), week 2 (S2), and week 4 (S3). First gingivitis and then plaque was scored. All gingivitis assessments were carried out by the same trained examiner (NLH). All plaque assessments were carried out by a second trained examiner (CEB). All examinations were carried out under the same conditions. All teeth were examined for both indices at six sites per tooth (disto-buccal, mid-buccal, mesio-buccal, disto-lingual, mid-lingual, mesio-lingual) except for 3rd molars. Criteria Gingivitis was assessed as the primary outcome using the bleeding on marginal probing index (BOMP) as described by Van der Weijden et al. (1994a, 1994b) and Lie et al. (1998). In short, the gingival margin is probed at an angle of approximately 60° to the longitudinal axis of the tooth and the absence or presence of bleeding is scored within 30 seconds of probing on a scale 0 - 2 (0 = no bleeding, 1 = pinprick bleeding, 2 = excessive bleeding). Plaque was assessed as a secondary outcome using the Turesky (1970) modification of the Quigley & Hein (1962) plaque index (TQHPI) as described in detail by Figure 2. OIP - oral irrigator with prototype tip Figure 1. Toothbrush - Oral-B indicator 35 Figure 3. OIS - oral irrigator with standard tip Figure 4. DF - standard waxed dental floss JIAP 10-005 Rosema.indd 4 04/01/2011 11:37:47 The Effect of Different Interdental Cleaning Devices on Gingival Bleeding 5 Flowchart Screening Screening 172 subjects Excluded 64 subjects Baseline (S1) Total: 108 subjects Group 1: 36 subjects Group 2: 36 subjects Group 3: 36 subjects BOMP & TQHPI Professional instruction 2 Weeks (S2) Drop-out 2 subjects Total: 106 subjects Group 1: 35 subjects Group 2: 35 subjects Group 3: 36 subjects BOMP & TQHPI Drop-out 2 subjects 4 Weeks (S3) Total: 104 subjects Group 1: 34 subjects Group 2: 34 subjects Group 3: 36 subjects BOMP & TQHPI Statistical Analyses (ITT) Total: 104 subjects Group 1: 34 subjects Group 2: 34 subjects Group 3: 36 subjects Figure 5. Flowchart Paraskevas et al. (2007). Briefly, the teeth were dyed using a new cotton swab with fresh disclosing solution (Mira-2-Ton®; Hager & Werken GmbH & Co. KG. Duisburg, Germany) for each quadrant in order to disclose the plaque. Subsequently, the absence or presence of plaque was recorded on a 6-point scale (0-5, 0 = no plaque, 5 = plaque covering more than two-thirds of the tooth surface). Study design This study was designed as single masked, 3-group parallel, 4-week home use experiment. After meeting the inclu- JIAP 10-005 Rosema.indd 5 sion criteria, completion of a medical questionnaire and informed consent, subjects returned to the clinic for their first (baseline) assessment (S1) for both clinical parameters (bleeding on marginal probing and plaque). At the start of the experiment all subjects received a unique trial number. Subjects were randomly assigned to one of three groups according to a randomization list (www.random. org). The allocation of products was carried out by the study coordinator, who was responsible for allocation concealment. All products were distributed in such a way that blindness of the examiners was assured. At the last visit (S3) the study coordinator assured blindness of the 04/01/2011 11:37:48 6 Journal of the International Academy of Periodontology (2011) 13/1 examiners by collecting the study products in a separate room from where the clinical examinations took place. Subjects were also instructed not to mention anything to the examiners that could lead to allocation disclosure. During the 4-week experimental phase OIS and OIP subjects used the OI once a day in the evening with lukewarm tap water and were instructed to finish one container of 500 ml at each occasion. Subjects in the control group (DF) used standard waxed dental floss once a day in the evening. At the baseline visit (S1), immediately following the baseline assessment, subjects used their allocated product for the first time. The study coordinator (NAMR) was present to provide detailed verbal instruction, a demonstration to ensure correct use, and aid with further personal instruction when necessary. Subjects in both OI groups were instructed to use the OI according to the instruction leaflet provided by the manufacturer. Subjects in the DF group were instructed to use their product according to the description of Van der Weijden et al. (2008). All subjects in each group were instructed to brush twice a day in their normal manner, once in the morning after breakfast and once in the evening. In the evening they subsequently used their assigned product (OI or DF). All participants were instructed to refrain from using any other oral hygiene product or device such as toothpicks, interdental brushes, mouthrinses, etc., during the study period. To check for compliance, subjects were asked to register the time of use of the products onto a calendar record chart. After two weeks (S2), subjects returned to the clinic for the second clinical assessment for both gingivitis and plaque. After four weeks (S3), subjects visited the clinic for their final assessment for both parameters. Subjects were asked to return all products provided for this study as well as the calendar record chart. On each occasion subjects were instructed to brush between 2 and 3 hours prior to their appointments to avoid the risk of increased bleeding on probing as a result of toothbrushing (Abbas et al., 1990). The day prior to each appointment all subjects received an SMS-message as a reminder with the following text: “Remember that you have an appointment at ACTA! Note that you need to brush your teeth 2-3 hours prior to your visit. See you tomorrow! ACTA.” After the final assessment habitual oral hygiene procedures were resumed. Data analyses The unit of analysis was the subject and collected data were analyzed as intention to treat. The bleeding scores were used as the main response variable (Galgut et al., 1998) and plaque scores as secondary response variable. A priori calculations with an alpha of 0.05, a difference of 0.0883 (between groups) of the bleeding index with 80% power, based on a pooled SD of 0.13 as derived JIAP 10-005 Rosema.indd 6 from previous studies supported a sample size of 105. An analysis of covariance (ANCOVA) with S1 as covariate and S3 as dependent variable was performed to compare groups over time (Heynderickx et al., 2005). Analyses comparing differences between the test and control groups at each time point were performed using non-parametric tests. Explorative analyses were performed to investigate the origin of the overall differences. P values of < 0.05 were accepted as statistically significant. Results Of 108 subjects who started the trial, four subjects did not complete the protocol. One chose not to continue the trial for personal reasons. Another left the country and moved abroad. Two did not attend the second visit because of scheduling conflicts. This resulted in a study population of 104 subjects providing evaluable data (Figure 5). The study population data on demographics and pre-study floss habits are presented in Table 1. No adverse events were reported by any of the subjects who participated in this study. Results for bleeding on probing are presented in Table 2. The overall ANCOVA analysis showed a statistically significant difference between the three groups (p = 0.007). Mean overall reductions after four weeks of use (S1 to S3) were 0.15 for the OIP group, 0.17 for the OIS group, and 0.02 for the DF group. The mean bleeding scores of the three groups did not differ significantly at baseline. At session 2 the scores decreased for all three groups. Post testing showed that both the OI groups provided significantly lower bleeding scores as compared to the DF group. At session 3 a statistically significant difference could be detected among the three groups. Post testing showed that again both the OI groups had significantly lower bleeding scores as compared to the DF group. The 95% confidence interval of the difference compared to the DF group at S3 was -0.27 ± -0.04 for the OIP group and -0.28 ± -0.05 for the OIS group. Results for plaque index are presented in Table 3. With regard to the plaque scores the overall ANCOVA analysis showed no statistically significant differences among the three groups (p = 0.126). Mean overall reductions after four weeks of use (S1 to S3) were -0.09 for the OIP group, 0.06 for the OIS group, and 0.01 for the DF group. Discussion Effective brushing remains the most obvious way of maintaining low levels of plaque and good gingival health. Gingivitis is known to be associated with the onset of periodontitis, and although the relationship between these two conditions may not be fully understood, the importance of maintaining good gingival health and 04/01/2011 11:37:48 The Effect of Different Interdental Cleaning Devices on Gingival Bleeding 7 Table 1. Demographic data and pre-study flossing habits of the study population. N Female Male Age [range] (SD) Daily floss users Weekly floss users Monthly floss users Seldom/never floss users Total OIP OIS DF 104 74 30 21.8 [18-36] 6 16 20 62 34 24 10 21.9 (3.2) 2 7 9 16 34 27 7 21.1 (2.3) 1 4 7 22 36 23 13 22.4 (3.1) 3 5 4 24 OIP, oral irrigation device with prototype jet tip; OIS, oral irrigation device with standard jet tip; DF, dental floss Table 2. Mean bleeding index (BOMP) and mean % bleeding scores for all groups at all sessions. N Session 1 Session 2 Session 3 OIP - index % 34 0.82 (0.25) 46 % 0.65 (0.24) 37 % 0.67 (0.26)† 39 % OIS index % 34 0.83 (0.23) 46 % 0.61 (0.27)* 34 % DF - index % 36 0.86 (0.26) 47 % 0.579 p - value (Kruskal Wallis) Relative Reduction Relative Reduction S1 – S2 S1 – S3 20 % 15 % 0.66 (0.26)* 38 % 26 % 17 % 0.74 (0.26) 41 % 0.84 (0.30) 47 % 13 % 0% 0.084 0.016 Standard deviation in parentheses. Univariate analyses of covariance with session 1 as covariate and session 3 as dependent variable. (p = 0.007). *Statistically significant difference compared to DF group, p < 0.05 (Mann-Whitney). †Statistically significant difference compared to DF group, p = 0.020 (Mann-Whitney). OIP, oral irrigation device with prototype jet tip; OIS, oral irrigation device with standard jet tip; DF, dental floss Table 3. Mean Quigley & Hein plaque scores ± standard deviation for all groups at all sessions. OIP OIS DF N Session 1 Session 2 Session 3 34 34 36 1.64 ± 0.43 1.79 ± 0.34 1.60 ± 0.26 1.61 ± 0.34 1.74 ± 0.29 1.51 ± 0.27 1.73 ± 0.37 1.73 ± 0.28 1.59 ± 0.27 Univariate analyses of covariance with session 1 as covariate and session 3 as dependent variable. (p = 0.126). OIP, oral irrigation device with prototype jet tip; OIS, oral irrigation device with standard jet tip; DF, dental floss preventing periodontitis is well recognised (Van Dyke et al., 1999). As the interproximal area is known as where the onset of gingival inflammation is likely to occur, the reason for interproximal plaque control seems clear. Although it is universally recognized that interproximal cleansing is essential for controlling periodontal disease (Löe, 1979), many people have difficulty accomplishing JIAP 10-005 Rosema.indd 7 this with traditional dental floss (Asadoorian, 2006). Thus, compliance with floss is low (Warren and Chater, 1996), and various adjuncts for interdental cleaning have been studied. Dental floss, toothpicks, woodsticks and interdental brushes have all been recommended for this purpose. The present study focussed on the ability to reduce gingival inflammation in a population of young individuals with moderate gingivitis using an OI. The OI works through the direct application of a pulsed stream of water or other solution. A study duration of four weeks was chosen to monitor the changes in the bleeding index, which meets the ADA guidelines on OI’s for studies assessing the effects of adjunctive therapies on reduction of gingivitis (ADA, 2008). Studies of longer duration will more clearly demonstrate the clinical benefit that subjects will obtain from this product. The efficacy of use of floss on the bleeding index was considered inconclusive in a systematic review by Berchier et al. (2008). The results of the present study are 04/01/2011 11:37:48 8 Journal of the International Academy of Periodontology (2011) 13/1 in support of this statement. In contrast, in the present study both OI groups did show statistically significant improvements after four weeks. At the end of the study both OI groups show a significant 15 - 17% reduction of the bleeding index as compared to baseline. For the DF group this difference was not observed. Comparisons among groups showed a significant difference at four weeks between the DF group and both OI groups. The absolute difference of 8% and 9% at four weeks for both OI groups as compared to the floss group reveals a relative effect of 17% (OIP) and 19% (OIS). In consideration of the ADA guidelines for oral irrigators, the results of the present study do not reach the lower limit of superiority of 20% as estimated proportionate reduction related to clinical relevance as compared to standard oral hygiene procedures (ADA, 2008). However, the ADA also has guidelines on adjunctive dental therapies (ADA, 1997). In those guidelines a lower limit of 15% is applied. The study outcomes of the present study do comply with this guideline, indicating a potential beneficial effect for the OI. With respect to plaque, the DF group started with a markedly lower score as compared to both OI groups. All subjects were instructed to brush 2-3 hours prior to examination, to reduce the risk of greater bleeding tendency (Abbas et al., 1990). As the difference in PI scores was consistent throughout the study and was not reflected in bleeding index scores, it seems that subjects who were randomly allocated to the floss group coincidently performed better instant plaque removal by brushing at visit days. In a study carried out by Galgut et al. (2000) the effect of unevenly distributed baseline data is discussed and it was concluded that this might not influence the results and the conclusions drawn. Historically, plaque reductions are considered a prerequisite for an oral hygiene device to be considered effective (Löe et al., 1965). A recent systematic review (Husseini et al., 2008) reported no statistically significant reduction in plaque when the OI was used as an adjunct to toothbrushing when compared to toothbrushing only. Despite a lack of effect on plaque index, the studies that were included in this review did find a significant effect on bleeding and gingival indices. The mechanisms of actions underlying these clinical changes for the bleeding index in the absence of a clear effect on plaque are not understood, although different hypotheses have been put forward (Husseini et al., 2008). One of the hypotheses is that supragingival irrigation alters the population of key pathogens, reducing gingival inflammation (Flemming et al., 1995). Another hypothesis is that the water-pulsation may alter the specific host-microbial interaction in the subgingival environment (Chaves et al., 1994). There is also the possibility that the beneficial action of an OI is at least partly because of the removal of loosely adherent soft deposits interfering with plaque JIAP 10-005 Rosema.indd 8 maturation and stimulation of the immune response (Frascella et al., 2000). Other explanations could be a mechanical stimulation of the gingiva or a combination of the above-mentioned factors (Frascella et al., 2000; Flemmig et al., 1990). Furthermore, irrigation may reduce the thickness of the plaque, which may not be easily detectable using 2-dimensional scoring systems (Jolkovsky et al., 1990). The absence of an effect for DF at four weeks may also seem surprising. A transient effect of 6% BI reduction was observed at two weeks. However, a recent systematic review supports this finding that dental floss has no significant effect on plaque or bleeding indices (Berchier et al., 2008). The small effect observed at two weeks is most likely the result of a novelty or Hawthorne effect. The Hawthorne effect is a reaction of subjects to the realization they are in a study and are being observed (Adair et al., 1984). The novelty effect and Hawthorne effect can be considered as certain placebo effects. The impact of a placebo effect should not be underestimated (Finniss et al., 2010). In a study by Feil et al. (2002), the Hawthorne effect was intentionally used and shown to improve oral health. The novelty effect is something that could have influenced all groups within this model. Subjects were pre-selected on having “no experience” with an OI, whereas only six out of the 104 were regular flossers (Table 1). The rebound that is observed from the 2-week to the 4-week follow-up is, however, most evident in the floss users. With respect to the Hawthorne effect, this is probably not only present in the DF group but also in both OI groups, as subjects were selected on having a bleeding index of > 50%. However at session 1 the bleeding index was already reduced to 46-47% for all three groups. This indicates that subjects already acted as if they were entered into the protocol before the first assessment of the primary response variable. The results of the present study add to the existing data and clearly show a reduction in inflammation from using an OI. Interestingly, the reduction in bleeding could not be linked to plaque removal. This is similar to data presented by Flemmig et al. (1990) showing no change in plaque scores for either the brushing group or the brushing and irrigation group from baseline to 6 months, but a significant difference in bleeding on probing and gingival index scores in favor of the irrigation group. Likewise, Flemmig et al. (1995) reported that the water irrigation group was significantly better at reducing bleeding on probing and gingival index scores compared to the regular oral hygiene group at six months. Also in this study there were no statistically significant differences detected in plaque scores among the groups. Chaves et al. (1994) found similar reductions in plaque scores for water irrigation compared to toothbrushing alone, and a significant difference for bleeding on probing in favor of the irrigation group at six months. 04/01/2011 11:37:48 The Effect of Different Interdental Cleaning Devices on Gingival Bleeding These studies support the present data in finding no correlation between reduction of plaque biofilm and inflammation in 3-6 months. Conclusion There is a long-standing, well-documented body of evidence supporting the use of an oral irrigator. An oral irrigator is at least as effective as dental floss for reducing gingival bleeding and gingivitis. When combined with manual toothbrushing the use of an oral irrigator, either with a prototype or standard jet tip, is significantly more effective in reducing gingival bleeding scores as compared to the use of dental floss, as determined within the limits of this 4-week study design. Acknowledgment The study was performed in commission of ACTA Research BV. Waterpik Inc, Fort Collins, CO, USA initiated the study project and provided study products. ACTA Research BV received financial support for their commitment to appoint this project to the Department of Periodontology of ACTA. D.M. Lyle is the director of professional and clinical affairs for Water Pik, Inc. The authors employed by ACTA declare that they have no conflict of interest. References ADA. Acceptance program guidelines: Adjunctive dental therapies for the reduction of plaque and gingivitis. American Dental Association Council on Scientific Affairs, September 1997. ADA. Acceptance program guidelines: Oral irrigating devices. American Dental Association Council on Scientific Affairs, 2008. Adair J.G. The Hawthorne effect: a reconsideration of the methodological artifact. Journal of Applied Psychology 1984; 69:334-345. Al-Mubarak, S., Ciancio, S., Aljada, A. et al. Comparative evaluation of adjunctive oral irrigation in diabetes. Journal of Clinical Periodontology 2002; 29:295-300. Asadoorian, J. Flossing. Canadian dental hygienists association position statement. Canadian Journal of Dental Hygiene 2006; 40:1-10. Abbas, F., Voss, S., Nijboer, A., Hart, A.A. and Van der Velden, U. The effect of mechanical oral hygiene procedures on bleeding on probing. Journal of Clinical Periodontology 1990; 17:199-203. Barnes, C.M., Russell, C.M., Reinhardt, R.A., Payne, J.B. and Lyle, D.M. Comparison of irrigation to floss as an adjunct to tooth brushing: effect on bleeding, gingivitis, and supragingival plaque. Journal of Clinical Dentistry 2005; 16:71-77. Berchier, C.E., Slot, D.E., Haps, S. and Van der Weijden, G.A. The efficacy of dental floss in addition to a toothbrush on plaque and parameters of gingival inflammation: a systematic review. International Journal of Dental Hygiene 2008; 6:265-279. Brown, L.J. and Löe, H. Prevalence, extent, severity and progression of periodontal disease. Periodontology 2000 1993; 2:57-71. Brownstein, C.N., Briggs, S.D., Schweitzer, K.L., Briner, W.W., and Kornman, K.S. Irrigation with chlorhexidine to resolve naturally occurring gingivitis. Journal of Clinical Periodontology 1990; 17:588-593. Burch, J.B., Lanese, R. and Ngam, P. A two-month study of the effects of oral irrigation and automatic toothbrush use in an adult orthodontic population with fixed appliances. American Journal of Orthodontics and Dentofacial Orthopedics 1994; 106:121-126. JIAP 10-005 Rosema.indd 9 9 Chaves, E.S., Kornman, K.S, Manwell, M.A., Jones, A.A., Newbold, D.A. and Wood, R.C. Mechanism of irrigation effects on gingivitis. Journal of Periodontology 1994; 65:1016-1021. Ciancio, S.G., Mather, M.L., Zambon, J.J. and Reynolds, H.S. Effect of chemotherapeutic agent delivered by an oral irrigation device on plaque, gingivitis, and subgingival microflora. Journal of Periodontology 1989; 60:310-315. Cobb, C.M., Rodgers, R.L. and Killoy, W.J. Ultrastructural examination of human periodontal pockets following the use of an oral irrigation device in vivo. Journal of Periodontology 1988; 59:155-163. Costerton J.W., Lewandowski Z., DeBeer D., Caldwell, D., Korber, D. and James, G. Biofilms, the customized microniche. Journal of Bacteriology 1994; 176:2137-2142. Cutler, C.W., Stanford, T.W., Abraham, C., Cederberg, R.A., Boardman, T.J. and Ross, C. Clinical benefits of oral irrigation for periodontitis are related to reduction of pro-inflammatory cytokine levels and plaque. Journal of Clinical Periodontology 2000; 27:134-143. Douglass, C.W. and Fox, C.H. (1993) Cross-sectional studies in periodontal disease: current status and implications for dental practice. Advances in Dental Research 1993; 7:25-31. Drisko, C.L., White, C.L., Killoy, W.J. and Mayberry, W.E. Comparison of dark-field microscopy and a flagella stain for monitoring the effect of a Water Pik on bacterial motility. Journal of Periodontology 1987; 58:381-386. Feil, P.H., Grauer, J.S., Gadbury-Amyot, C.C., Kula, K. and McCunniff, M.D. Intentional use of the Hawthorne effect to improve oral hygiene compliance in orthodontic patients. Journal of Dental Education 2002; 66:1129-1135. Felo, A., Shibly, O., Ciancio, S.G., Lauciello, F.R. and Ho, A. Effects of subgingival chlorhexidine irrigation on peri-implant maintenance. American Journal of Dentistry 1997; 10:107-110. Fine, J.B., Harper, D.S., Gordon, J.M., Hovliaras, C.A. and Charles, C.H. Journal of Periodontology 1994; 65:30-36. Finniss, D.G., Kaptchuk, T.J., Miller, F. and Benedetti, F. Biological, clinical, and ethical advances of placebo effects. Lancet 2010; 375:686-695. Flemmig, T.F., Newman, M.G., Doherty, F.M., Grossman, E., Meckel, A.H. and Bakdash, M.B. Supragingival irrigation with 0.06% chlorhexidine in naturally occurring gingivitis. I. 6 month clinical observations. Journal of Periodontology 1990; 61:112-117. Flemmig, T.F., Epp, B., Funkenhauser, Z., et al. Adjunctive supragingival irrigation with acetylsalicylic acid in periodontal supportive therapy. Journal of Clinical Periodontology 1995; 22:427-433. Frascella J.A., Fernández P., Gilbert R.D. and Cugini M. A randomized, clinical evaluation of the safety and efficacy of a novel oral irrigator. Am J Dent 2000; 13:55–58. Galgut, P.N. and O’Mullane, D. Statistical analysis of data derived from clinical variables of plaque and gingivitis. Journal of Clinical Periodontology 1998; 7:549-553. Galgut, P.N. Management of data used in clinical trials which is unevenly distributed at baseline. Current Medical Research Opinion 2000: 16:46-55. Gorur, A., Lyle, D.M., Schaudinn, C. and Costerton, J.W. Biofilm removal with a dental water jet. Compendium of Continuing Education in Dentistry 2009; 30:1-6. Heynderickx, I. and Engel, J. Statistical methods for testing plaque removal efficacy in clinical trials. Journal of Clinical Periodontology 2005; 32:677-683. Husseini, A., Slot, D.E. and Van der Weijden, G.A. The efficacy of oral irrigation in addition to a toothbrush on plaque and the clinical parameters of periodontal inflammation: a systematic review. International Journal of Dental Hygiene 2008; 6:304-314. Jepsen, S. The role of manual toothbrushes in effective plaque control: Advantages and limitations. In: Lang, N.P., Attström, R. and Löe, H. (Eds): Proceedings of the European Workshop on Mechanical Plaque Control. Berlin: Quintessenz Verlag 1998, 121-137. 04/01/2011 11:37:48 10 Journal of the International Academy of Periodontology (2011) 13/1 Jolkovsky, D.L., Waki, M.Y., Newman, M.G., et al. Clinical and microbiological effects of subgingival and gingival marginal irrigation with chlorhexidine gluconate. Journal of Periodontology 1990; 61:663-669. Lie, M.A., Timmerman, M.F., Van der Velden, U. and Van der Weijden, G.A. Evaluation of 2 methods to assess gingival bleeding in smokers and non-smokers in natural and experimental gingivitis. Journal of Clinical Periodontology 1998; 25:695-700. Löe, H., Theilade, E. and Jensen, S.B. Experimental gingivitis in man. Journal of Periodontology 1965; 36:177-187. Löe, H. Mechanical and chemical control of dental plaque. Journal of Clinical Periodontology 1979; 6:32-36. Lobene, R.R. The effect of a pulsed water pressure cleansing device on oral health. Journal of Periodontology 1969; 40:667-670. Marsh, P.D. and Bradshaw, D.J. Dental plaque as a biofilm. Journal of Industrial Microbiology 1995; 15:169-175. Newman, M.G., Cattabriga, M., Etienne, D., et al. Effectiveness of adjunctive irrigation in early periodontitis: multi-center evaluation. Journal of Periodontology 1994; 65:224-229. Quigley, G.A. and Hein, J.W. Comparative cleansing efficiency of manual and power brushing. Journal of the American Dental Association 1962; 65:26-29. Paraskevas, S, Rosema, N.A., Versteeg, P., Timmerman, M.F., van der Velden, U. and Van der Weijden G.A. The additional effect of a dentifrice on the instant efficacy of toothbrushing: a crossover study. Journal of Periodontology 2007; 78:1011-1016. Research, Science and Therapy Committee, American Academy of Periodontology Position Paper. The role of supra- and subgingival irrigation in the treatment of periodontal diseases. Journal of Periodontology 2005; 76:2015-2027. Research, Science and Therapy Committee, American Academy of Periodontology Position Paper. Treatment of plaque-induced gingivitis, chronic periodontitis, and other clinical conditions. Journal of Periodontology 2001; 72:1790-1800. Sharma, N.C., Lyle, D.M., Qaqish, J.G., Galustians, J. and Schuller, R. Effect of a dental water jet with orthodontic tip on plaque and bleeding in adolescent patients with fixed orthodontic appliances. American Journal of Orthodontics and Dentofacial Orthopedics 2008; 133:565-571. JIAP 10-005 Rosema.indd 10 Slot, D.E., Dörfer, C.E. and Van der Weijden GA. The efficacy of dental floss in addition to a toothbrush on plaque and parameters of gingival inflammation: a systematic review. International Journal of Dental Hygiene 2008; 6:265-279. Turesky, S., Gilmore, N.D. and Glickman, L. Reduced formation by chloromethyl analogue of vitamin C. Journal of Periodontology 1970; 41:41-43. Van Dyke, T.E., Offenbacher, S., Pihlstrom, B., Putt, M.S. and Trummel, C. What is gingivitis? Current understanding of prevention, treatment, measurement, pathogenesis and relation to periodontitis. Journal of the International Academy of Periodontology 1999; 1:3-15. Van der Weijden, G.A., Timmerman, M.F., Nijboer, A., Lie, M.A. and Van der Velden, U. A comparative study of electric toothbrushes for the effectiveness of plaque removal in relation to toothbrushing duration. Timer study. Journal of Clinical Periodontology 1993; 20:476-481. Van der Weijden, G.A., Timmerman, M.F., Saxton, C.A., Russell, J.I., Huntington, E. and Van der Velden, U. Intra-/inter-examiner reproducibility study of gingival bleeding. Journal of Periodontal Research 1994a; 29:236-241. Van der Weijden, G.A., Timmerman, M.F., Reijerse, E., Nijboer, A. and Van der Velden, U. Comparison of different approaches to assess bleeding on probing as indicators of gingivitis. Journal of Clinical Periodontology 1994b; 21:589-594. Van der Weijden, G.A., Echevaria, J.J., Sanz, M. and Lindhe J. Mechanical supragingival plaque control. In: Lindhe, J., Lang, N.P. and Karring, T. (Eds): Clinical Periodontology and Implant Dentistry, 5th edition. Munskgaard. Wiley-Blackwell, 2008. Walsh, T.F., Glenwright, H.D. and Hull, P.S. Clinical effects of pulsed oral irrigation with 0.02% chlorhexidine digluconate in patients with adult periodontitis. Journal of Clinical Periodontology 1992; 19:245-248. Warren, P.R. and Chater, B.V. An overview of established interdental cleaning methods. Journal of Clinical Dentistry 1996; 7:65-69. 04/01/2011 11:37:48 Journal of the International Academy of Periodontology 2011 13/1: 11–16 Additive or Synergistic Antimicrobial Effects of Amoxicillin and Metronidazole on Whole Plaque Samples: A Preliminary Report Clemens Walter1,2, Eva M. Kulik3, Roland Weiger1, Nicola U. Zitzmann1 and Tuomas Waltimo3 Department of Periodontology, Endodontology and Cariology, School of Dentistry, University Basel, Switzerland; 2Department of Oral Surgery, School of Dentistry, University of Birmingham, United Kingdom; 3Institute for Preventive Dentistry and Oral Microbiology, School of Dentistry, University Basel, Switzerland 1 Abstract Objective: In vitro data on the susceptibility of oral bacteria to the combination of metronidazole and amoxicillin is limited. The aim of this preliminary study was to determine the susceptibility of whole subgingival plaque samples to amoxicillin and metronidazole and to their combination. Methods: Prior to any treatment procedures subgingival plaque samples from patients with severe generalized periodontitis were taken. Appropriate dilutions were plated on Columbia blood agar supplemented with the following agents: 3 µg/mL amoxicillin, 8 µg/mL amoxicillin, 8 µg/mL metronidazole, 16 µg/mL metronidazole, 3 µg/mL amoxicillin plus 8 µg/mL metronidazole or 8 µg/mL amoxicillin plus 16 µg/mL metronidazole. All plates were incubated anaerobically at 36°C for 14 days and the colony forming units (CFU) were determined. Results: Both applied metronidazole concentrations were able to decrease the CFU counts by approximately one order of magnitude in a log10 scale. Amoxicillin 3 µg/mL revealed a reduction of 2.4 log10 CFU, whereas 50% of the samples did not grow on the plates supplemented with 8 µg/mL of amoxicillin. There was no anaerobic bacterial growth on agar plates supplemented with the combination of amoxicillin and metronidazole even at the lower antibiotic concentrations. Conclusion: Susceptibility screening of subgingival samples to metronidazole and amoxicillin and to their combination seems to offer a rational basis for the selection of adjunctive antibiotic therapy Key words: Antibiotics, synergistic effect, aggressive periodontitis, metronidazole, amoxicillin Introduction Periodontal diseases are multifactorial biofilm-associated infections. A distinct differentiation between aggressive and chronic forms is difficult (Meyer et al., 2004), even on the basis of microbiological findings (Mombelli et al., 2002; Ximenez-Fyvie et al., 2006; Schacher et al., 2007). Hence, the diagnosis of “aggressive periodontitis” is primarily based on clinical and radiological characteristics, on patient’s age, and on findings derived during clinical follow-up. Due to the infection-induced nature of periodontal diseases, antimicrobial therapies based on microbiological examinations may improve the treat- Correspondence to: Prof. Dr. Nicola U. Zitzmann, Department of Periodontology, Endodontology and Cariology, School of Dentistry, University Basel, Hebelstrasse 3, CH4056 Basel, Switzerland. Email: n.zitzmann@unibas.ch ment outcome of advanced and/or aggressive forms of periodontitis. First attempts to control periodontal diseases with the adjunctive use of antibiotics included systemic administration of tetracyclines, amoxicillin with or without clavulanic acid, clindamycin and metronidazole (Listgarten et al., 1978; Lekovic et al., 1983; Gordon et al., 1985; Magnusson et al., 1989). Another adjunctive treatment approach was topical administration of various antibiotics or antiseptics (Lindhe et al., 1979; Needleman and Watts, 1989; Stabholz et al., 2000). Two decades ago, the combination of metronidazole and amoxicillin - so called “van Winkelhoff-Cocktail” - was introduced as an adjunctive systemic therapy for periodontitis treatment (Van Winkelhoff et al., 1989). This regimen was specifically designed for treatment of diseases associated with Aggregatibacter (Actinobaccillus) actinomycetemcomitans, for which a synergistic in vitro effect between the two substances or their metabolites has been reported © International Academy of Periodontology JIAP 10-004 Zitzmann.indd 11 04/01/2011 11:39:02 12 Journal of the International Academy of Periodontology (2011) 13/1 (Pavicic et al., 1994a; Pavicic et al., 1994b). Clinical studies on aggressive forms of periodontitis have revealed improved outcomes within observation periods up to five years, provided that the adjunctive treatment with the combination of amoxicillin and metronidazole was strictly combined with mechanical biofilm removal (Buchmann et al., 2002; Guerrero et al., 2005; Kaner et al., 2007a; Kaner et al., 2007b). Moreover, improved clinical advantages of this regimen were found in a placebocontrolled study comparing the antibiotic combination to the agents alone, again as adjunctive to mechanical, non-surgical periodontal treatment (Rooney et al., 2002). In this report, the treatment outcomes of subjects with advanced chronic periodontal disease were independent from the initial microbiological findings. Recently, this strategy of combining amoxicillin and metronidazole was used for the treatment of generalized “aggressive periodontitis” without targeting against specific microorganisms (Guerrero et al., 2005). Whenever antibiotics are administrated as an adjunctive periodontal treatment, existing or possibly developing resistance of the associated microflora should be carefully considered. In vitro findings have suggested that there are remarkable differences in resistance profiles of certain oral bacterial species (Van Winkelhoff et al., 2005; Lakhssassi et al., 2005). Recent findings in microbiological susceptibility testing have indicated the rationale of the examination of mixed microbial cultures instead of, or in addition to, the individual disease-associated strains (Karbach et al., 2007). Such in vitro data about bacterial susceptibility to the combination of amoxicillin and metronidazole is hitherto scarce. The aim of the present preliminary study was to determine the susceptibility of whole subgingival plaque samples to amoxicillin and metronidazole and to their combination. Materials and methods Patients and sampling Four generally healthy patients with severe generalized chronic or aggressive periodontitis were recruited from the pool of patients from the Department of Periodontology, Endodontology and Cariology at the School of Dental Medicine, University of Basel, Switzerland. Diagnosis was based on clinical and radiographic findings, related to age and the severity of destruction (Table 1, Figure 1a-b, Armitage, 1999). Clinical measurements of probing pocket depth and attachment level were performed with the probe PCPUNC-15 (Hu-Friedy, Chicago, IL, USA). All recruited patients (one female and three males with a mean age of 40.8 years) were current or former heavy smokers and had neither received any earlier periodontal treatment nor systemic or topical antibiotics one year prior to the sampling. The female patient was not pregnant. JIAP 10-004 Zitzmann.indd 12 Subgingival plaque samples were taken for antibiotic resistance analysis. At least the two deepest periodontal pockets with bleeding on probing were selected for microbiological sampling. Supragingival plaque was removed, the sampling site was isolated using cotton rolls and gently dried with air. A sterile paper point was inserted to the bottom of the pocket, left in place for 20 s and placed in 0.5 ml of thioglycolate broth (bioMérieux, Genf, Switzerland; Casas et al., 2007). Microbiological procedures Immediately after sampling, pooled paper points were vortexed for one minute and serially diluted in thioglycolate broth. For the determination of the total anaerobic bacterial count, 100 mL of the dilutions were plated on Columbia blood agar plates (Columbia Agar Base [BBL Becton Dickinson, Allschwil, Switzerland] enriched with 4 mg/L hemin, 1 mg/L menadione, and 50 ml/L human blood). For quantification of the proportion of microorganisms resistant to either amoxicillin and/or metronidazole, Columbia blood agar plates supplemented with the following concentrations of the respective antimicrobial agent were used: 3 mg/mL amoxicillin (Fluka, Buchs, Switzerland), 8 mg/mL amoxicillin, 8 mg/mL metronidazole (Fluka), 16 mg/mL metronidazole, 3 mg/mL amoxicillin plus 8 mg/mL metronidazole or 8 mg/mL amoxicillin plus 16 mg/mL metronidazole. The concentrations of the antibiotics were adopted from van Winkelhoff et al. (2000) and/or the Clinical Laboratory and Standards Institute (2007). All plates were incubated anaerobically (10% CO2, 10% H2, 80% N2) at 36°C for 14 days and the colony forming units (CFUs) were determined. Results Microbial findings Microbiological data are presented in Table 2. The total anaerobic plaque count (CFU) ranged from 3.1 x 106 to 7.2 x 107 among the plaque samples, and the percentage of black-pigmented bacteria ranged from 40 to 80%. All samples showed a decrease of bacterial growth on agar by approximately 1 log with both concentrations of the antibiotic agent (8 mg/mL and 16 mg/mL). All agar plates supplemented with 3 mg/ mL amoxicillin showed a reduced bacterial growth by log 2.4, whereas two out of four samples revealed no growth on the plates supplemented with 8 mg/mL of amoxicillin (Table 2). On agar plates supplemented with the combination of amoxicillin and metronidazole, no anaerobic bacterial growth was detected even at lower antibiotic concentrations. 04/01/2011 11:39:02 Additive and/or synergistic in vitro effect of antibiotics 13 Table 1. Profile of study patients and clinical characteristics. Patient Age Periodontal diagnosis Smoking status Number of teeth Number of sites with PPD Number of ≥ 6 mm sites with BOP+ 1 45 GAgP Former smoker 30 pack years 29 165 174 2 32 GAgP Current smoker 15 pack years 29 92 133 3 38 GAgP Current smoker 17 pack years 25 69 96 4 48 GChP Current smoker 30 pack years 27 25 46 BOP, bleeding on probing; GAgP, generalized aggressive periodontitis; GChP, generalized chronic periodontitis Figure 1a. Figure 1b. Figure 1. Patient N° 2 was diagnosed with generalized aggressive periodontitis due to extensive bone loss at the age of 32 years. a) Clinical intraoral photographs; b) Full-mouth periapical radiographs Table 2. Microbiological characteristics and results of the antibiotic susceptibility analyses. Bacterial growth on Bacterial growth on agar plates supplemented with different concentrations of metronidazole or amoxicillin agar plates (control) Patient 1 2 3 4 CFU % BPB 4.0 x 107 1.0 x 107 3.1 x 106 7.2 x 107 40 50 40 80 CFU CFU CFU CFU CFU CFU amoxicillin amoxicillin metronida- metronida- amoxicillin 3 mg/mL amoxicillin 8 mg/mL + metronidazole +metronidazole zole zole 8 mg/mL 3 mg/mL 16 mg/mL 8 mg/mL 16 mg/mL 8 mg/mL 9.0 x 104 8.0 x 103 8.4 x 104 4.0 x 105 1.0 x 104 3.0 x 105 6.0 x 106 6.0 x 105 6.0 x 105 1.3 x 106 1.5 x 107 6.0 x 105 1.3 x 106 1.5 x 106 - - BPB, black-pigmented bacteria; CFU, colony forming units JIAP 10-004 Zitzmann.indd 13 04/01/2011 11:39:04 14 Journal of the International Academy of Periodontology (2011) 13/1 Discussion The present preliminary study using subgingival plaque samples demonstrated reduced bacterial growth in the presence of low concentrations of metronidazole or amoxicillin, while higher amoxicillin concentrations inhibited bacterial growth in two out of four samples. Interestingly, the combination of metronidazole and amoxicillin was effective against microorganisms in all subgingival plaque samples at lower antibiotic concentrations. This in vitro observation suggests an additive or synergistic mode of action for these agents, which is likely to be beneficial for infection control, as demonstrated by recent clinical studies (van Winkelhoff et al., 1989; Buchmann et al., 2002; Rooney et al., 2002; Guerrero et al., 2005; Kaner et al., 2007a; Kaner et al., 2007b). It may be hypothesized that the targeted use of this additive/synergistic effect, which is either based on growth inhibition or on bacteriocidal effects, may offer a strategy against the development and/or the control of resistant strains. The introduced method testing microbial susceptibility to a frequently administrated combination of antibiotics is a novel approach, which enlightens the capacity of additive and/or synergistic effects between the two substances. A synergistic effect of two antibiotics needs to be evaluated on a species level, and was documented for Aggregatibacter actinomycetemcomitans (Pavicic et al., 1994a; Pavicic et al., 1994b). The authors suggested a higher rate of metronidazole uptake by bacterial cells simultaneously incubated with amoxicillin. Resistance of anaerobic bacteria to metronidazole hardly ever occurred (Seifert and Dalhoff, 2010). In the current material, bacterial growth was detected in all four subgingival plaque samples, which is indicative of metronidazole-resistant strains and emphasizes the need for susceptibility testing in selected patients with infections involving anaerobic bacteria. The results of the current study should be, however, interpreted with caution due to the limited number of subjects included, and the lack of specific bacterial strain characterisation. However, the mixed subgingival plaque samples used here represented the expected general characteristics in terms of relative proportions of back-pigmented anaerobes in the total culturable flora. This preliminary study was restricted to current or former heavy smokers, who have an increased risk for the onset and progression of periodontal diseases (Warnakulasuriya et al., 2010). Cigarette smoking is likely to affect the composition of the oral microflora due to a decrease in oxygen tension in periodontal pockets, and may promotes a selection of anaerobic bacteria (Hanioka et al., 2000). However, the literature has been indecisive as to whether a specific smoking-associated microbial profile exists (van Winkelhoff et al., 2001; van der Velden et al., 2003). Interestingly, recent evidence JIAP 10-004 Zitzmann.indd 14 from a randomized controlled trial suggests a benefit of adjunctive antimicrobial therapy with metronidazole and amoxicillin in the non-surgical periodontal treatment of smokers with chronic periodontitis (Matarazzo et al., 2008). The culture technique used in the current investigation may have some shortcomings: (i) restricted to growth of viable bacteria, (ii) strict sampling and transport conditions essential, (iii) specific laboratory equipment and experienced personnel required for bacterial culturing, (iv) time needed for bacterial growth on appropriate media, (v) specific pathogens in the subgingival plaque may not be detected. However, the main advantage of the technique used is the probability of an analysis of bacterial resistance against the combination of antibiotics, in particular against amoxicillin and metronidazole. The diversity of the oral microflora, reaching up to 700 different bacterial species (Kazor et al., 2003), makes it impossible to analyze every single bacterial strain regarding a genetic profile encoding for antibiotic resistance. In addition, the molecular mechanisms of bacterial resistance to antibiotics are quite far from being completely understood. Therefore, the antibiotic susceptibility of a subgingival plaque sample or of putative periodontal pathogens needs to be analyzed by conventional culture techniques (Armitage, 2003). A major concern of the presented approach is the natural biofilm association of the subgingival bacterial samples analysed. A biofilm is a difficult therapeutic target because of its three-dimensional structure, which protects the bacteria from the host response as well as from antimicrobial agents (Socransky and Haffajee, 2002; Eick and Pfister, 2004). The methodology of the present report allowed the interactions between culturable microorganisms, but no attempt was made to mimic other characteristics of the subgingival plaque. Different results may be expected when a biofilm of mixed microbial samples is formed on an appropriate substrate prior to their susceptibility testing. However, such an approach is currently not available. The chosen methodology aims to provide an approach for clinically relevant susceptibility testing. According to the contemporary understanding of the pathogenesis, periodontal diseases are caused by an opportunistic infection with a conglomerate of potentially periopathogenic microorganisms organized in the subgingival biofilm. A number of different test methods and procedures are available for qualitative and quantitative microbiological diagnostics of putative periopathogens. However, the pathogenic potential of a certain putative periodontal pathogen against the host can hitherto not be determined. Moreover, major individual differences in the immune response are caused by a number of acquired or genetic factors. Although specific bacteria have a periopathogenic potential or 04/01/2011 11:39:05 Additive and/or synergistic in vitro effect of antibiotics may initiate periodontal inflammation, it is still difficult to determine the microbiota responsible for the onset and progression of disease in the individual subject. Thus, in the diagnosis and therapy of periodontal diseases, microbiological identification and susceptibility testing of single disease-associated strains may be of limited value (Mombelli et al., 2002; Sanz et al., 2004). Instead or in addition to the conventional approach of microbiological diagnostics, susceptibility testing of the entire subgingival plaque sample may offer additional valuable information for the choice of the antibiotic to be administered adjunctively. Improved therapy outcomes indicate that patients with periodontal diseases - particularly those with highly destructive forms (aggressive and/or advanced) - may profit from an adjunctive antibiotic therapy using amoxicillin and metronidazole (Guerrero et al., 2005; Kaner et al., 2007a). However, due to the increased use of antibiotics and the alarming development of resistant strains, antibiotics should be administered with care, and testing the susceptibility of a given individual’s microflora may have an increasing importance (Walker, 1996; Van Winkelhoff et al. 2005; Lakhssassi et al., 2005; Walter and Weiger, 2006). Therefore, microbial testing can not be recommended for routine dental practise. However, some patients, in particular those in need of adjunctive antimicrobial therapy, may profit from the information about potential therapeutic targets (Armitage, 2003). Susceptibility testing of whole subgingival samples to metronidazole and amoxicillin and to their combination seems to offer a rational diagnostic tool to the selection of adjunctive antibiotic therapy. In the event of an unfavorable response, i.e. bacterial growth on agar plates supplemented with amoxicillin and metronidazole, another antibiotic has to be tested and subsequently applied for adjunctive antimicrobial therapy. The current report about susceptibility analyses of subgingival plaque samples was initiated as a proof-ofprinciple study. The microbial results derived from the audit of four cases may indicate a potential benefit for further analysis in a larger clinical microbiological trial. Acknowledgements We gratefully acknowledge the technical assistance of Mrs. Krystyna Lenkeit (Dental School, University Basel, Switzerland) and the constructive criticism of Prof. em. Jürg Meyer (Dental School, University Basel, Switzerland). There is no conflict of interest. References Armitage, G. C. Development of a classification system for periodontal diseases and conditions. Annuals of Periodontology 1999; 4:1-6. Armitage, G. C. Diagnosis of periodontal diseases. Journal of Periodontology 2003; 74:1237-1247. Buchmann, R., Nunn, M. E., Van Dyke, T. E. and Lange, D. E. Aggressive periodontitis: 5-year follow-up of treatment. Journal of Periodontology 2002; 73:675-683. JIAP 10-004 Zitzmann.indd 15 15 Casas, A., Herrera, D., Martin-Carnes, J., Gonzalez, I., O’Connor, A. and Sanz, M. Influence of sampling strategy on microbiologic results before and after periodontal treatment. Journal of Periodontology 2007; 78:1103-1112. Clinical and Laboratory Standards Institute Methods for antimicrobial susceptibility testing of anaerobic bacteria; approved standard M11-A7. CLSI, Payne, WA, USA. 2007. Eick, S. and Pfister, W. Efficacy of antibiotics against periodontopathogenic bacteria within epithelial cells: an in vitro study. Journal of Periodontology 2004; 75:1327-1334. Gordon, J., Walker, C., Lamster, I. et al. Efficacy of clindamycin hydrochloride in refractory periodontitis. 12-month results. Journal of Periodontology 1985; 56:75-80. Guerrero, A., Griffiths, G. S., Nibali, L. et al. Adjunctive benefits of systemic amoxicillin and metronidazole in non-surgical treatment of generalized aggressive periodontitis: a randomized placebo-controlled clinical trial. Journal of Clinical Periodontology 2005; 32:1096-1107. Hanioka, T., Tanaka, M., Takaya, K., Matsumori, Y. and Shizukuishi, S. Pocket oxygen tension in smokers and non-smokers with periodontal disease. Journal of Periodontology 2000; 71:550-554. Kaner, D., Christan, C., Dietrich, T., Bernimoulin, J. P., Kleber, B. M. and Friedmann, A. Timing affects the clinical outcome of adjunctive systemic antibiotic therapy for generalized aggressive periodontitis. Journal of Periodontology 2007a; 78:1201-1208. Kaner, D., Bernimoulin, J. P., Hopfenmüller, W., Kleber, B. M. and Friedmann, A. Controlled-delivery chlorhexidine chip versus amoxicillin/metronidazole as adjunctive antimicrobial therapy for generalized aggressive periodontitis: a randomized controlled clinical trial. Journal of Clinical Periodontology 2007b; 34:880-891. Karbach, J., Callaway, A., Willershausen, B., Wagner, W., Geibel, M. A. and Al-Nawas, B. Antibiotic resistance testing of the total implant-associated micro-flora and its pure isolates. European Journal of Medical Research 2007; 12:120-128. Kazor, C. E., Mitchell, P. M., Lee, A. M., Stokes, L. N., Loesche, W. J., Dewhirst, F. E. and Paster, B. J. Diversity of bacterial populations on the tongue dorsa of patients with halitosis and healthy patients. Journal of Clinical Microbiology 2003; 41:558-563. Lakhssassi, N., Elhajoui, N., Lodter, J. P., Pineill, J. L. and Sixou, M. Antimicrobial susceptibility variation of 50 anaerobic periopathogens in aggressive periodontitis: an interindividual variability study. Oral Microbiology and Immunology 2005; 20:244-252. Lekovic, V., Kenney, E. B., Carranza, F.A. Jr. and Endres, B. The effect of metronidazole on human periodontal disease. A clinical and bacteriological study. Journal of Periodontology 1983; 54:476-480. Lindhe, J., Heijl, L., Goodson, J. M. and Socransky, S. S. Local tetracycline delivery using hollow fiber devices in periodontal therapy. Journal of Clinical Periodontology 1979; 6:141-149. Listgarten, M. A., Lindhe, J. and Hellden, L. Effect of tetracycline and/or scaling on human periodontal disease. Clinical, microbiological, and histological observations. Journal of Clinical Periodontology 1978; 5:246-271. Magnusson, I., Clark, W. B., Low, S. B., Maruniak, J., Marks, R. G. and Walker, C.B. Effect of non-surgical periodontal therapy combined with adjunctive antibiotics in subjects with “refractory” periodontal disease. (I). Clinical results. Journal of Clinical Periodontology 1989; 16:647-653. Matarazzo, F., Figueiredo, L. C., Cruz, S. E. B. , Faveri, M. and Feres, M. Clinical and microbiological benefits of systemic metronidazole and amoxicillin in the treatment of smokers with chronic periodontitis: a randomized placebo-controlled study. Journal of Clinical Periodontology 2008; 35:885–896. Meyer, J., Lallam-Laroye, C. and Dridi, M. Aggressive periodontitis what exactly is it? Journal Clinical Periodontology 2004; 31:586-587. Mombelli, A., Casagni, F. and Madianos, P. N. Can presence or absence of periodontal pathogens distinguish between subjects with chronic and aggressive periodontitis? A systematic review. Journal of Clinical Periodontology 2002; 29 Suppl 3:10-21. 04/01/2011 11:39:05 16 Journal of the International Academy of Periodontology (2011) 13/1 Needleman, I. G. and Watts, T.L. The effect of 1% metronidazole gel in routine maintenance of persistent furcation involvement in human beings. Journal of Periodontology 1989; 60:699-703. Pavicic, M. J., van Winkelhoff, A. J., Pavicic-Temming, Y. A. and de Graaff, J. Amoxycillin causes an enhanced uptake of metronidazole in Actinobacillus actinomycetemcomitans: a mechanism of synergy. Journal of Antimicrobial Chemotherapy 1994a; 34:1047-1050. Pavicic, M. J., van Winkelhoff, A. J., Douque, N. H., Steures, R. W. and de Graaff, J. Microbiological and clinical effects of metronidazole and amoxicillin in Actinobacillus actinomycetemcomitans-associated periodontitis. A 2-year evaluation. Journal of Clinical Periodontology 1994b; 21:107-112. Rooney, J., Wade, W. G., Sprague, S. V., Newcombe, R. G. and Addy, M. Adjunctive effects to non-surgical periodontal therapy of systemic metronidazole and amoxycillin alone and combined. A placebo-controlled study. Journal of Clinical Periodontology 2002; 29:342-350. Sanz, M., Lau, L., Herrera, D., Morillo, J. M. and Silva, A. Methods of detection of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythensis in periodontal microbiology, with special emphasis on advanced molecular techniques: a review. Journal of Clinical Periodontology 2004; 31:1034-1047. Schacher, B., Baron, F., Rossberg, M., Wohlfeil, M., Arndt, R. and Eickholz, P. Aggregatibacter actinomycetemcomitans as indicator for aggressive periodontitis by two analysing strategies. Journal of Clinical Periodontology 2007; 34:566-573. Seifert, H. and Dalhoff, A. German multicentre survey of the antibiotic susceptibility of Bacteroides fragilis group and Prevotella species isolated from intra-abdominal infections: results from the PRISMA study. Journal of Antimicrobial Chemotherapy 2010; 65:2405-2410 Socransky, S. S. and Haffajee, A. D. Dental biofilms: difficult therapeutic targets. Periodontology 2000 2002; 28:12-55. Stabholz, A., Shapira, L., Mahler, D. et al. Using the PerioChip in treating adult periodontitis: an interim report. Compendium of Continuing Education in Dentistry 2000; 21:325-8, 330-332. JIAP 10-004 Zitzmann.indd 16 van Winkelhoff, A. J., Rodenburg, J. P., Goene, R. J., Abbas, F., Winkel, E. G. and de Graaff, J. Metronidazole plus amoxycillin in the treatment of Actinobacillus actinomycetemcomitans-associated periodontitis. Journal of Clinical Periodontology 1989; 16:128-131. van Winkelhoff, A. J., Gonzales, D. H., Winkel, E. G., DellemijnKippuw, N., Vandenbroucke-Grauls, C. M. and Sanz, M. Antimicrobial resistance in the subgingival microflora in patients with adult periodontitis. A comparison between The Netherlands and Spain. Journal of Clinical Periodontology 2000; 27:79-86. van Winkelhoff, A. J., Bosch-Tijhof, C. J., Winkel, E. G. and van der Reijden, W. A. Smoking affects the subgingival microflora in periodontitis. Journal of Periodontology 2001; 72:666-671. van Winkelhoff, A. J., Herrera, D., Oteo, A. and Sanz, M. Antimicrobial profiles of periodontal pathogens isolated from periodontitis patients in The Netherlands and Spain. Journal of Clinical Periodontology 2005; 32:893-898. van der Velden, U., Varoufaki, A., Hutter, J. W., et al. Effect of smoking and periodontal treatment on the subgingival microflora. Journal of Clinical Periodontology 2003; 30:603-610. Walker, C. B. The acquisition of antibiotic resistance in the periodontal microflora. Periodontology 2000 1996; 10:79-88. Walter, C. and Weiger, R. Antibiotics as the only therapy of untreated chronic periodontitis: a critical commentary. Journal of Clinical Periodontology 2006; 33:938-939. Warnakulasuriya, S., Dietrich, T., Bornstein, M. M., et al. Oral health risks of tobacco use and effects of cessation. International Dental Journal 2010; 60:7-30 Ximenez-Fyvie, L. A., Maguer-Flores, A., Jacobo-Soto, V., LaraCordoba, M., Moreno-Borjas, J. Y. and Cantara-Maruri, E. Subgingival microbiota of periodontally untreated Mexican subjects with generalized aggressive periodontitis. Journal of Clinical Periodontology 2006; 33:869-877. 04/01/2011 11:39:05 Journal of the International Academy of Periodontology 2011 13/1: 17–26 Clinical and Microbiological Comparison of Three Non-surgical Protocols for the Initial Treatment of Chronic Periodontitis Carlos Serrano, Nidia Torres, Angela Bejarano, Marcela Caviedes and María Eugenia Castellanos School of Dentistry, Pontificia Universidad Javeriana, Bogotá, Colombia Abstract Objective: To compare the clinical and microbiological effects of three protocols for nonsurgical periodontal therapy, including full-mouth scaling and root planing plus systemic antibiotics, on the treatment of chronic periodontitis patients. Methods: Twenty-nine patients diagnosed with moderate to severe chronic periodontitis, selected according to specific criteria, were randomly assigned to one of three treatment groups: quadrant scaling, full-mouth scaling, and full-mouth scaling supplemented by systemic antibiotics. Antibiotic selection was based on the results of individual susceptibility testing. Oral hygiene instructions and reinforcement were given during the study. All patients received a clinical periodontal and microbiological examination at baseline and at reexamination, 4-6 weeks after therapy. Means and standard deviations were calculated and differences between groups were analyzed via the Kruskal-Wallis test, p < 0.05. Results: The mean age of the study sample was 49.1 ± 11.6 years old, and there were 17 men and 12 women. Patients treated with antibiotics showed antimicrobial susceptibility for amoxicillin and doxycycline. All study groups showed a similar significant improvement in periodontal parameters. Plaque scores were reduced in a range of 29.0% to 42.6%. Bleeding on probing was reduced by 34.8% to 55.0%; the reduction for the full-mouth scaling group was larger. Mean reduction in pocket depth was 1.2 to 1.3 mm in all groups. Mean bacterial counts were reduced in the groups receiving full-mouth treatment, but not in the quadrant treatment group. Conclusion: The three protocols for non-surgical periodontal treatment demonstrated a similar positive effect on clinical parameters; however, only full-mouth treatment groups showed a reduction in anaerobic microbial counts at re-examination. Key words: Chronic periodontitis, scaling and root planing, antibiotics, full-mouth scaling Introduction The main goal of periodontal therapy is to control the infection associated with chronically inflamed tissues through a series of activities aimed at reducing the bacterial destructive effect, such as oral hygiene instruction, subgingival debridement and surgical pocket reduction. These protective measures, when reinforced by meticulous self-performed oral hygiene and regular professional maintenance, lead to the re-establishment of periodontal health (Tunkel et al., 2002; van der Wejden and Timmerman, 2002). Initial therapy for disease includes root surface instrumentation procedures, scaling and root planing, usually performed on jaw quadrants Correspondence to: Carlos Serrano, MSc., Specialist in Periodontology, Department of Periodontal System, School of Dentistry, Pontificia Universidad Javeriana, Carrera 7 # 40-62, Edificio de Odontología, Bogotá, Colombia. E-mail: serrano-c@ javeriana.edu.co during a series of appointments. Systematic reviews on non-surgical periodontal therapy have considered scaling and root planing an effective treatment, measured by clinical parameters such as reduction of bleeding upon probing, reduction of probing pocket depth and gain in probing attachment level (van der Wejden and Timmerman, 2002; Hung and Douglass, 2002; Hallmon and Rees, 2003). Studies on the microbiological effect of scaling and root planing have shown that it produces a marked disruption of the subgingival biofilm, leading to decreased levels and proportion of sites colonized by periodontal pathogens (Teles et al., 2006). Considering that periodontal pathogens can be found in different intra-oral niches besides periodontal pockets, such as the tongue, the tonsils, the saliva and other mucous membranes (van Winkelhoff et al., 1986; Asikainen et al., 1991), and that translocation of pathogens from the above niches or untreated periodontal pockets to recently treated pockets is possible (Quyrinen et al., 1996), the Leuven University research group de- © International Academy of Periodontology JIAP 10-003 Serrano.indd 17 22/12/2010 09:41:22 18 Journal of the International Academy of Periodontology (2011) 13/1 veloped the full-mouth disinfection concept (Quyrinen et al., 1995). This treatment protocol includes scaling and root planing in two sessions within 24 hours, and usage of different therapeutic forms of clorhexidine for brushing the tongue, rinsing the mouth, spraying the tonsils and irrigating pockets. Reports from the original research group have concluded that full-mouth disinfection has a beneficial effect in the treatment of moderate and severe periodontitis, evidenced by 15% greater reduction in bleeding on probing, 1.3-1.8 mm greater reduction in probing pocket depth, and ≥ 1.5 mm additional gain in probing attachment level over quadrant scaling (Mongardini et al., 1999; Quyrinen et al., 2006). A modification of the full-mouth disinfection protocol is one stage full-dentition scaling and root planing only, since it was identified as the key component responsible for the additional clinical and microbiological improvements over traditional treatment (Quyrinen et al., 2000). During recent years, several clinical reports on the effect of full-mouth treatment concepts have yielded contradictory findings (Apatzidou and Kinane, 2004; Koshy et al., 2005; Wennstrom et al., 2005; JervoeStorm et al., 2006; Zanatta et al., 2006). Two recent systematic reviews concluded that differences between full-mouth disinfection, full-mouth scaling and root planing and traditional treatment were modest, so that any of the three treatment modalities may be used for initial therapy for chronic periodontitis (Lang et al., 2008; Eberhard et al., 2008). Considering the lack of significant differences between full-mouth treatment concepts and traditional therapy referred to in systematic reviews (Lang et al., 2008; Eberhard et al., 2008), and the additional clinical benefit over scaling and root planing described for the adjunctive use of systemic antibiotics (Herrera et al., 2002; Haffajee et al., 2003), Cionca et al. (2009) studied the effects of combining metronidazole and amoxicillin, proper oral hygiene, and scaling and root planing over 24 hours in chronic periodontitis patients. The authors conducted a placebo-controlled clinical trial of six months duration, reporting significant clinical improvements for both patient groups, but greater reduction in the number of pockets > 4 mm and percentage of sites with bleeding on probing for subjects receiving amoxicillin and metronidazole. In conclusion, the authors stated that systemic antibiotics significantly improved the clinical outcome of full-mouth non-surgical periodontal therapy and reduced the need for additional therapy. The purpose of this study was to compare the clinical and microbiological effects of three protocols for non-surgical periodontal therapy, including full-mouth scaling and root planing plus systemic antibiotics, on the treatment of chronic periodontitis patients. JIAP 10-003 Serrano.indd 18 Materials and Methods Study design This was a parallel design, randomized clinical trial performed at the Specialist Periodontal Clinic, School of Dentistry, Javeriana University in Bogotá, Colombia. Approval for the protocol was obtained from the Ethics Committee, Javeriana University; all patients signed an informed consent form before the start of the study. Patients Twenty-nine patients were recruited from new referrals to the Javeriana University, Specialist Clinic, according to the following criteria: age ranging from 25-75 years; possession of a minimum of 16 teeth; diagnosis of chronic periodontitis (according to the criteria of the International Classification System for Periodontal Diseases [Armitage, 1999]) with severity ranging from moderate to severe and multiple sites showing clinical attachment loss ≥ 3 mm in all jaw quadrants; available for participation in a follow-up examination after two months. Patients were excluded if they had uncontrolled systemic conditions, such as diabetes or cardio-vascular disease, or required antibiotic coverage for treatment; were pregnant or breast-feeding; were taking medication that could affect the periodontal condition or had received antibiotic therapy during the previous three months; had suspected or confirmed allergy to antibiotic agents, including β-lactam or tetracycline group antibiotics; or were current smokers, defined as having a consumption rate of ≥ 10 cigarettes per day. Experimental design Enrolled subjects were randomly assigned by a computer-generated table to one of three non-surgical treatment modalities (Research Randomizer, Social Psychology Network, 1997-2009): Control group (Q): scaling and root planing quadrant by quadrant at weekly intervals. Full-mouth scaling and root planing (FM): fullmouth scaling and root planing performed over two consecutive days. Full-mouth scaling and root planing plus antibiotics (FMa): full-mouth scaling and root planing performed over two consecutive days combined with the administration of systemic antibiotics determined by susceptibility testing for each subject. All patients had a clinical examination and microbiological sampling before any treatment procedure was performed (baseline), and at a period 4-6 weeks after the last appointment for scaling and root planing (re-examination). 22/12/2010 09:41:22 Clinical and Microbiological Comparison of Three Non-surgical Protocols for the Initial Treatment of Chronic Periodontitis Clinical examination Clinical measurements of bleeding on probing, probing pocket depth and clinical attachment level were performed on all present teeth at six tooth surfaces. Four surfaces were examined for the O´Leary plaque score [percentage of presence/absence of plaque using a disclosing solution: mesial, buccal, distal and lingual (O´Leary, 1972)]. Probing pocket depth (PPD) was measured with a manual periodontal probe to the closest millimeter. Bleeding on probing (BoP) was recorded as the percentage of sites bleeding after probing pocket depth measurements. Specific analysis was performed for moderate pockets (5 - 6 mm), and for deep pockets (≥ 7 mm). The gingival margin (GM) level was measured from the gingival margin to a reference point, either the cemento-enamel junction or the margin of a restoration. In case of gingival recession, the gingival margin level has a negative value. The probing attachment level (PAL) was measured as ± GM – PD. Reduction in the percentage of sites with PPD ≥ 4 mm from baseline to re-examination was recorded. Microbiological examination After cotton roll isolation and removal of supragingival plaque, pooled microbial samples were taken with paper cones from the deepest periodontal pocket in each quadrant for 15 seconds (Mombelli et al., 1991). Samples were placed in vials containing anaerobic VMGA III transport medium (Möller, 1966) and transferred for processing. At the microbiological laboratory, samples were vortex-mixed for 30 seconds and serially diluted 10-fold five times. One hundred μL of 10-1 dilution was plated on blood agar and incubated for three days under aerobic conditions. One hundred μL of 10-4 dilution was plated on anaerobic Wilkins-Chalgren agar (Oxoid, United Kingdom); plates were incubated for seven days at 37 ºC in 80% N2, 10% CO2 and 10% H2. Colony forming units (CFU) were enumerated on blood and Wilkins-Chalgren agar plates after the period of incubation. Presumptive identification of periodontal pathogens was performed based on colony morphology, Gram staining, aerotolerance test and a commercial identification micromethod system (RapID ANA II, Oxoid, United Kingdom). Patients randomly assigned to the FMa group had an antimicrobial susceptibility testing on identified periodontal pathogen colonies, mainly Porphyromona gingivalis, Prevotella intermedia, Fusobacterium nucleatum and Wollinela spp. Five different commercially available antibiotic strips were used: amoxicillin, metronidazole, azithromycin, tetracycline and doxycycline (E-test, AB Biodisk, Solna, Sweden). Viable pure colonies were homogenized in 0.85% saline solution and adjusted to the MacFarland turbidity standard 1.0. Using a sterile glass rod, 0.1 mL of the inoculum was spread over Wilkins-Chalgren agar plates and allowed to dry for 15 minutes. E-test strips JIAP 10-003 Serrano.indd 19 19 were gently placed over the agar surface and incubated under anaerobic conditions for three days. The intersection between the zone of bacterial inhibition and the E-test strip represented the minimal inhibitory concentration (MIC). The antibiotic with the lowest MIC was selected to be used; if several tests were made for the same patient the antibiotic with the lowest average MIC was chosen. Treatment procedures Scaling and root planing were performed by three periodontal graduate students (AB, MC and MEC). The FM and FMa received scaling and root planing on two consecutive days; in addition, for the FMa group the antibiotic showing the lowest minimal inhibitory concentration was prescribed for a one-week period. The control group (Q) received scaling and root planing in four sessions, quadrant by quadrant, at one-week intervals. All scaling was performed using manual curettes (Hu-Friedy, Chicago, IL, U.S.A.) and ultrasonic magnetostrictive instruments (Cavitron, Dentsply, York, PA, U.S.A.) without any time restriction. At the baseline examination, all patients were given detailed oral hygiene instructions, including toothbrush and dental floss or inter-dental brush use. No mouth rinses were used during the study. Patients received oral hygiene reinforcement at weeks 2 and 4. Teeth judged ready to be extracted due to poor periodontal condition were extracted and not included for analysis; restorations with overhangs were replaced during treatment. Statistical analysis Mean and standard deviation values for the O´Leary plaque score, percentage of BoP, PPD, PAL and percentage of pockets were calculated for each subject. Mean and standard deviation of logarithmically transformed CFU counts were calculated. The changes within groups from baseline to re-examination were analyzed using the paired Wilcoxon test. The differences between groups were analyzed using the Kruskal-Wallis test. The level of significance was set at p < 0.05. Results A total of twenty-nine patients completed the clinical study: 10 patients in the control Q group, nine patients in the FM group, and 10 patients in the FMa group. The mean age of the patient sample was 49.1 ± 11.6 years old; there were 17 men and 12 women. Twenty-six patients were non-smokers, two patients were smoking 2 - 5 cigarettes per day, and a third patient smoked 2 - 3 cigarettes per week. Demographic characteristics of the patient sample are described in Table 1. Antimicrobial susceptibility testing results for the 10 subjects included in the FMa group revealed that in four subjects, the lowest MIC value was for amoxicil- 22/12/2010 09:41:22 20 Journal of the International Academy of Periodontology (2011) 13/1 Table 1. Demographic characteristics of the patient sample Number of patients Mean age Gender male/female Q FM FMa 10 46.6 (10.9) 6/4 9 52.8 (10.6) 6/3 10 48.2 (13.3) 4/6 0 0 20 20 40 40 % 60 60 80 80 100 100 Q, control; FM, full mouth scaling and root planing; FMa, full mouth scaling and root planing plus systemic antibiotics Q FM BL Q FMa FM BL Final Figure 2. Percentage of sites with bleeding on probing 0 20 40 60 80 Figure 1. Change in O`Leary plaque score FMa Final Q FM BL FMa Final Figure 3. Percentage of sites with probing pocket depth (PPD) ≥ 4 mm JIAP 10-003 Serrano.indd 20 22/12/2010 09:41:23 Clinical and Microbiological Comparison of Three Non-surgical Protocols for the Initial Treatment of Chronic Periodontitis lin; for the other six subjects the lowest value was for doxycycline. As a consequence, four subjects received amoxicillin 875 mg twice a day for seven days; and six subjects received doxycycline 100 mg once a day for seven days. Plaque scores All study groups showed a high level of plaque accumulation at baseline before oral hygiene instructions were given, Q 56.7% ± 22.8, FM 77.8% ± 38.7 and FMa 47.3% ± 18.2. Nevertheless, with a random group assignment the O`Leary plaque score was higher for the FM group than for the other two groups. At re-examination, all three groups showed a statistically significant improvement in the O`Leary plaque score, with mean reduction values ranging from 29.0% for the FMa group to 42.6% for the FM; no statistically significant differences were detected among the three groups at re-examination (Figure 1). Bleeding on probing Percentage of sites bleeding on probing was equally high for all three groups at baseline: at re-examination, a statistically significant reduction in percentage of sites bleeding on probing was present in all groups. For the Q, the BoP percentage decreased from 78.7 ± 26.3% to 44.0 ± 29.0%, for the FM from 88.6 ± 17.8% to 33.6 ± 24.0%, and for the FMa from 80.0 ± 14.5% to 46.3 ± 27.9%. There were no statistically significant differences when the three groups were compared; however, the FM group obtained a 20% greater BoP percentage reduction when compared with the other two groups (Figure 2). Reduction in the percentage of pockets At baseline, the percentage of sites with increased PPD varied from 36.3% ± 20.9 for the FMa group, to 45.0% ± 28.1 for the Q group. A statistically significant reduction in the percentage of sites showing PPD ≥ 4 mm was observed at re-examination compared with baseline in all three groups. The reduction in percentage had a range from 12 to 17%; the greatest reduction was obtained in the Q group, and the least in the FM group. However, no statistically significant differences were obtained between groups (Figure 3). 21 Probing pocket depth The mean baseline PPD was similar in all three groups: it varied from 5.0 ± 0.6 mm for the Q group to 5.5 ± 0.7 mm for the FM group. At re-examination, all three groups showed a 1.2 - 1.3 mm significant reduction of PPD; no significant differences were present among the groups. When a separate analysis for pockets initially 5 - 6 mm and ≥ 7 mm was performed, no significant differences were found for mean change values among the three groups, but greater variability in the healing response of deep pockets was found for the Q group (Table 2, Figures 4, 5). Not all patients exhibited ≥ 7 mm deep pockets at baseline: three patients in the Q group and one patient each in the FM and FMa groups did not have pockets in that depth category. Probing attachment level A significant gain in PAL of about 1.0 mm was measured at re-examination for the three groups. No significant differences were found among the groups. Microbiological anaerobic CFU counts Great variability in log-transformed anaerobic CFU counts characterized the subjects in the three groups, which was reflected in high standard deviation values. As a partial consequence, no significant differences were observed in CFU counts at re-examination compared with baseline. A decrease in log-transformed CFU counts was measured for the FM and FMa groups, 0.34 and 1.33 respectively. In contrast, a slight increase was measured for the Q group, -0.41 log-transformed CFU count. Nevertheless, no significant differences were obtained among the three study groups (Figure 6). Of interest, only in the FMa group were there patients whose oral flora was below microbiological culture detection levels for anaerobes at re-examination (Figure 6). Discussion The results of the present study showed that periodontal therapy in all three study groups resulted in a significant improvement of periodontal clinical parameters. The percentage of BoP sites was reduced in the range of 34 - 55%, PPD was reduced by 1.2 - 1.3 mm, and the PAL gain was approximately 1 mm. These changes in Table 2. Mean probing pocket depth (SD) values according to 5 - 6 and ≥ 7 mm depth categories Q Baseline Re-examination Change FM FMa 5-6 ≥7 5-6 ≥7 5-6 ≥7 5.2 (0.1) 4.0 (0.6) 1.2 (0.5) 7.4 (0.3) 5.3 (1.6) 2.1 (1.3) 5.5 (0.2) 4.2 (0.9) 1.3 (0.6) 8.0 (0.8) 6.2 (0.8) 1.8 (0.9) 5.4 (0.2) 4.0 (0.7) 1.4 (0.5) 7.5 (0.4) 5.8 (1.1) 1.7 (0.9) Q, control; FM, full mouth scaling and root planing; FMa, full mouth scaling and root planing plus systemic antibiotics JIAP 10-003 Serrano.indd 21 22/12/2010 09:41:23 Journal of the International Academy of Periodontology (2011) 13/1 3 4 Milímetros 5 6 22 Q FM BL FMa Final 2 4 Milímetros 6 8 10 Figure 4. Probing pocket depth reduction from baseline: 5 - 6 mm depth category Q FM BL FMa Final 0 2 4 6 Figure 5. Probing pocket depth reduction from baseline: ≥ 7 mm depth category Q FM BL FMa Final Figure 6. Log-transformed anaerobic bacterial colony counts JIAP 10-003 Serrano.indd 22 22/12/2010 09:41:23 Clinical and Microbiological Comparison of Three Non-surgical Protocols for the Initial Treatment of Chronic Periodontitis clinical parameters are in the same range as results of systematic literature reviews on the effects of nonsurgical periodontal therapy (van der Wejden and Timmerman, 2002; Hung and Douglass, 2002). The majority of subjects in the present study showed great severity of periodontal disease, as over 40% of the surfaces were affected by periodontal pockets with a mean PPD range of 5.0 - 5.5 mm, and a large number of pockets ≥ 7 mm deep were present. No significant differences were found between the control group receiving quadrant scaling and root planing, and the other two groups receiving full-mouth scaling and root planing. This is in agreement with two recently published systematic reviews on the effects of full-mouth treatment concepts: they concluded that only modest differences were present between quadrant and full-mouth treatment (Eberhard et al., 2008; Lang et al., 2008). Noteworthy, in the present study the FM group had a 20% greater reduction in BoP percentage compared with the other two groups; however, due to a large standard deviation, this difference was not statistically significant. The systematic review by Lang et al. (2008) reported an opposite finding, as the weighted mean difference for BoP percentage reduction was 8.45% larger in the quadrant scaling groups. Of interest, no additional clinical benefit was obtained from systemic antibiotic administration even if the antibiotic showing the lowest microbial inhibitory concentration was selected. This is contradictory to the conclusion of systematic reviews on the effects of systemic antimicrobials as adjuncts to non-surgical periodontal treatment. The review of Herrera et al. (2002) reported a small additional gain in PAL of 0.3 mm and additional reduction of PPD ranging from 0.05 to 0.6 mm for patients receiving systemic antibiotics. Specific significant additional effects were found for the use of spiramycin regarding PPD change, and amoxicillin plus metronidazole regarding PAL change. The review of Haffajee et al. (2003) found that adjunctive systemic antibiotics led to an additional gain in PAL of 0.29 mm for pockets initially 4 - 6 mm deep, and of 0.45 mm for pockets initially ≥ 7 mm deep. Separate analysis by antibiotic type showed that tetracycline and metronidazole had a significant adjunctive benefit, followed by borderline results for the combination of amoxicillin plus metronidazole. In the present study, no greater reduction of PPD or gain of PAL was found in patients receiving systemic antibiotics, even when considering specific categories of initial PPD. As a possible explanation for the lack of any additional effect for systemic antibiotic administration, patient selection or antibiotic regime could be analyzed. The current sample age range was wide (25 - 75 years old), which could lead to the inclusion of subjects with different rates of disease progression and tissue destruction, even if all patients exhibited JIAP 10-003 Serrano.indd 23 23 clinical characteristics of chronic periodontitis. The 10 patients included in the FMa group were distributed to receive amoxicillin (four cases) or doxycycline (six cases); this reduced number of subjects does not allow analyzing the effect of each antibiotic in addition to non-surgical treatment. The combination of different antibiotics was not tested in the present study, even if the combination of amoxicillin and metronidazole in addition to scaling and root planing has been found to improve the clinical outcome of periodontal therapy on different populations (Pavicic et al., 1994; Winkel et al., 2001; Guerrero et al., 2005). The combination was not used for the present population due to the large levels of antimicrobial resistance to metronidazole. It may be that antibiotic regimens could have different adjunctive clinical benefits based on population variation for microbial antibiotic susceptibility. Nevertheless, healing in pockets initially ≥ 7 mm deep demonstrated a smaller range of variability in the FMa than in the Q group, where box plots for baseline and re-examination nearly overlap. The less variable healing response in the antibiotic group could lead to clinical differences during a longer healing time. Several methods of describing the effect of periodontal therapy on microbiological parameters have been used in the literature: for example, change in bacterial counts, proportion of colonized sites, or level of specific periodontal pathogens, including presence or absence at different threshold values (Teles, 2006). The systematic review by Lang et al. (2008) stated an equally positive improvement in microbiological parameters for quadrant and full-mouth scaling and root planing. However, in the present study no improvement in bacterial counts was seen in the Q group, whereas both full-mouth treatment groups showed decreased bacterial counts, the change being larger in the group that received antibiotics. Using similar methods, Quyrinen et al. (2000) found a small decrease in microbial counts for quadrant scaling, compared to a larger 1 logarithmic unit decrease in full-mouth treatment protocols, a comparable finding to the present study. On the contrary, studies that have used polymerase chain reaction techniques to detect specific periodontal pathogens before and after periodontal treatment have not found significant differences between quadrant and full-mouth treatment groups (Jervoe-Storm et al., 2007; Koshy et al., 2005). An exception occurred for Treponema denticola, which had a larger reduction in the full-mouth treatment group in the report by Apatzidou et al. (2004). The articles by Cionca et al. (2009, 2010) performed a clinical and microbiological comparison between fullmouth scaling and full-mouth scaling supplemented by systemic amoxicillin and metronidazole. The clinical results showed a significant clinical improvement for both treatment groups: bleeding on probing was re- 22/12/2010 09:41:23 24 Journal of the International Academy of Periodontology (2011) 13/1 duced by 37 to 45%, PPD was reduced by 1.2 - 1.3 mm, and gain in PAL was 0.7 to 0.9 mm., showing a similar clinical improvement to the present study. The main outcome of therapy in the Cionca et al. (2009) report was the number of sites with PPD ≥ 4 mm and bleeding on probing. This figure was significantly reduced in both treatment groups, with a statistically greater reduction for the group receiving antibiotics: 4.4 versus 1.3 remaining bleeding pockets. The present study used a different method, as the percentage of sites ≥ 4 mm deep, independent of bleeding on probing, was calculated. The microbiological study by Cionca et al. (2010) reported lower bacterial counts for both study groups, without any significant difference between them, similar to the present study. The addition of systemic antibiotics produced a greater reduction in the detection frequency of Aggregatibacter actinomycetemcomitans and P. gingivalis. Significant levels of antibiotic resistance to periodontal pathogens in samples taken from Colombian patients has led to the suggestion of performing antibiotic susceptibility testing before adjunctive antibiotic therapy (Ardila et al., 2010; Serrano et al., 2009). In the present study, amoxicillin or doxycycline was used for the treatment of the 10 patients included in the FMa group. Few studies have analyzed the effect of amoxicillin administration alone as an adjunct to nonsurgical periodontal therapy, while the combination of amoxicillin and metronidazole is more common (van Winkelhoff et al., 1992). It has been reported that the combination regimen produced a larger improvement in clinical parameters compared to placebo therapy, especially for patients who were initially positive for A. actinomycetemcomitans and P. gingivalis, and who became negative at re-examination (Pavicic et al., 1994; Winkel et al., 2001). However, a combination regimen was not used in the present study, as bacterial isolates showed significant resistance against metronidazole, a finding that has been described by other research groups in the same geographic area (Jaramillo et al., 2005; Ardila et al., 2010). The use of systemic doxycycline, 100 mg a day, as an adjunct of periodontal therapy, has produced additional benefits in the periodontal treatment of diabetic patients (Grossi et al., 1997; Martorelli de Lima et al., 2004; Llambés et al., 2005). Lately, those prescribing doxycycline have looked not only for its antimicrobial effect but also for inhibition of matrix metalloproteinases (Salvi and Lang 2005). An inclusion criterion in systematic reviews about the effects of full-mouth non-surgical periodontal therapy has been study duration of 3 or 6 months (Eberhardt et al., 2008; Lang et al., 2008). Nevertheless, in the present study re-examination was performed after 4 to 6 weeks in order to assess the need for surgical periodontal therapy in patients affected primarily by severe chronic periodontitis. A consensus report by the American Academy JIAP 10-003 Serrano.indd 24 of Periodontology stated that a 4- to 6-week interval seems appropriate to assess the initial response to nonsurgical periodontal therapy (Ciancio, 1989). The review article by Segelnick and Weinberg (2006) proposed to perform re-evaluation of basic periodontal therapy 4 to 8 weeks after scaling and root planing, based on evidence from clinical studies and the histology of periodontal wound healing. However, Badersten et al. (1981, 1984) reported that clinical improvement after therapy leveled off after 3 - 4 months for moderate periodontal cases, but the periodontal condition could continue to improve even for 6 to 9 months in advanced periodontal cases. The present study showed a trend toward larger PPD reduction for deep pockets and greater microbial count reduction in the group receiving full-mouth therapy supplemented by antibiotics; probably this trend would become significant in a longer healing time. In the present study, scaling and root planing was performed by periodontal graduate students. Different literature reports showed that experienced operators have better results regarding calculus removal during non-surgical instrumentation than graduate students (Brayer et al., 1989; Fleischer et al., 1989). Nevertheless, results of the present study are similar to the outcome of non-surgical therapy reported in systematic reviews in the literature. Conclusion The three protocols for non-surgical periodontal treatment demonstrated a similar positive effect on clinical parameters; however, only full-mouth treatment groups showed a reduction in anaerobic microbial counts at re-examination. Acknowledgement and conflict of interest The authors declare there are no conflicts of interest. This study was supported by a research grant from the Research Promotion Office, Javeriana University. References Apatzidou, D.A. and Kinane, D.F. Quadrant root planing versus same-day full-mouth root planing I. Clinical findings. Journal of Clinical Periodontology 2004; 31:132-140. Apatzidou, D.A., Riggio, M.P. and Kinane, D.F. Quadrant root planing versus same-day full-mouth root planing II: Microbiological findings. Journal of Clinical Periodontology 2004; 31:141-148. Ardila, C.M., Granada, M.I. and Guzman, I.C. Antibiotic resistance of subgingival species in chronic periodontitis patients. Journal of Periodontal Research 2010; 45:557-563. Armitage G.C. Development of a classification system for periodontal diseases and conditions. Annals of Periodontology 1999; 4:1-6. Asikainen, S., Alaluusua, S. and Saxén, L. Recovery of A. actinomycetemcomitans from teeth, tongue and saliva. Journal of Periodontology 1991; 62:203-206. Badersten, A., Nilveus, R. and Egelverg, J. Effect of non-surgical periodontal therapy. I. Moderately advanced periodontitis. Journal of Clinical Periodontology 1981; 8:57-72. 22/12/2010 09:41:23 Clinical and Microbiological Comparison of Three Non-surgical Protocols for the Initial Treatment of Chronic Periodontitis Badersten, A., Nilveus, R. and Egelberg, J. Effect of non-surgical periodontal therapy. II. Severely advanced periodontitis. Journal of Clinical Periodontology 1984; 11:63-76. Brayer, W.K., Mellonig, J.T., Dunlap, R.L., Marinak, K.W. and Carson, R.E. Scaling and root planing effectiveness: the effect of root surface access and operator experience. Journal of Periodontology 1989; 60:67-72. Ciancio, S.G. Non-surgical periodontal treatment. In: Proceeding of the World Workshop in Clinical Periodontics. American Academy of Periodontology; 1989: 11-14 Cionca, N., Giannopoulou, C., Ugolotti, G. and Mombelli, A. Amoxicillin and metronidazole as an adjunct to full-mouth scaling and root planing of chronic periodontitis. Journal of Periodontology 2009; 80:364-371. Cionca, N., Giannopoulou, C., Ugolotti, G. and Mombelli, A. Microbiologic testing and outcomes of full-mouth scaling and root planing with or without amoxicillin/metronidazole in chronic periodontitis. Journal of Periodontology 2010:81:15-23. Eberhardt, J., Jervoe-Storm, P.M., Needleman, I., Worthington, H. and Jepsen, S. Full-mouth treatment concepts for chronic periodontitis: a systematic review. Journal of Clinical Periodontology 2008; 35:591-604. Fleischer, H.C., Mellonig, J.T., Brayer, W.K., Gray, J.L. and Barnett, J.D. Scaling and root planing efficacy in multirooted teeth. Journal of Periodontology 1989; 60:402-409. Grossi, S.G., Skrepcinski, F.B., deCaro, T., et al. Treatment of periodontal disease in diabetes reduces glycated hemoglobin. Journal of Periodontology 1997; 68:713-719. Guerrero, A., Griffiths, G.S., Nibali, L., et al. Adjunctive benefit of systemic amoxicillin and metronidazole in non-surgical treatment of generalized aggressive periodontitis: a randomized placebo-controlled clinical trial. Journal of Clinical Periodontology 2005; 32:1096-1107. Haffajee, A.D., Socransky, S.S. and Gunsolley, J.C. Systemic antiinfective periodontal therapy. A systematic review. Annals of Periodontology 2003; 8:115-181. Hallmon, W.W. and Rees, TD. Local anti-infective therapy: mechanical and physical approaches. A systematic review. Annals of Periodontology 2003; 8:99-114. Herrera, D., Sanz, M., Jepsen, S., Needleman, I. and Roldán, S. A systematic review on the effect of systemic antimicrobials as an adjunct to scaling and root planing in periodontitis patients. Journal of Clinical Periodontology 2002; 29(Suppl. 3):136-159. Jaramillo, A., Arce, R.M., Herrera, D., Betancourth, M., Botero J.E. and Contreras, A. Clinical and microbiological characterization of periodontal abscesses. Journal of Clinical Periodontology 2005; 32:1213-1218. Jervoe-Storm, P.M., Semaan, E., AlAhdab, H., Engel, S., Fimmers, R. and Jepsen, S. Clinical outcomes of quadrant root planing versus full-mouth root planing. Journal of Clinical Periodontology 2006; 33:209-215. Jervoe-Storm, P.M., AlAhdab, H., Semaan, E., Fimmers, R. and Jepsen, S. Microbiological outcome of quadrant versus fullmouth root planing as monitored by real-time PCR. Journal of Clinical Periodontology 2007; 34:156-163. Koshy, G., Kawashima, Y., Kiji, M., et al. Effects of single-visit fullmouth ultrasonic debridement versus quadrant-wise ultrasonic debridement. Journal of Clinical Periodontology 2005; 32:734-743. Lang, N.P., Tan, W.C., Krähenmann, M.A. and Zwahlen, M. A systematic review of the effects of full-mouth debridement with and without antiseptics in patients with chronic periodontitis. Journal of Clinical Periodontology 2008; 35(Suppl. 8):8-21. Llambés, F., Silvestre, F.J., Hernández-Mijares, A., Guiha, R. and Caffesse, R. Effect of non-surgical periodontal treatment with or without doxycycline on the periodontium of type 1 diabetic patients. Journal of Clinical Periodontology 2005; 32:915-920. Martorelli de Lima, A.F., Cury, C.C., Palioto, D.B., Duro, A.M., Da JIAP 10-003 Serrano.indd 25 25 Silva, R.C. and Wolff, L.F. Therapy with adjunctive doxycycline local delivery in patients with type 1 diabetes mellitus and periodontitis. Journal of Clinical Periodontology 2004; 31:648-653. Möller, AJ. Microbiological examination of root canals and periapical tissues of human teeth. Methodological studies. Odontologisk Tidskrift 1966; 74(Suppl. 1):1-380. Mombelli, A., McNabb, H. and Lang, NP. Black-pigmenting Gramnegative bacteria in periodontal disease. II. Screening strategies for detection of P. gingivalis. Journal of Periodontal Research 1991; 26:308-313. Mongardini, C., van Steenberghe, D., Dekeyser, C. and Quirynen, M. One stage full-mouth versus partial-mouth disinfection in the treatment of chronic adult or generalized early-onset periodontitis. I. Long-term clinical observations. Journal of Periodontology 1999; 70:632-645. O’Leary, T.J., Drake, R.B. and Naylor, J.E. The plaque control record. Journal of Periodontology 1972; 43:38. Pavicic, M.J., van Winkelhoff, A.J., Douqué, N.H., Steures, R.W. and de Graaf, J. Microbiological and clinical effect of metronidazole and amoxicillin in Actinobacillus actinomycetemcomitans-associated periodontitis. A 2-year evaluation. Journal of Clinical Periodontology 1994; 21:107-112. Quirynen, M., Bollen, C.M., Vandekerckhove, B.N., Dekeyser, C., Papaioannou, W. and Eyssen, H. Full- versus partial-mouth disinfection in the treatment of periodontal infections: shortterm clinical and microbiological observations. Journal of Dental Research 1995; 74:1459-1467. Quirynen, M., Papaioannou, W. and van Steenberghe, D. Intraoral transmission and the colonization of oral hard surfaces. Journal of Periodontology 1996; 23:960-970. Quirynen, M., Mongardini, C., Pauwels, M., Bollén, C., van Eldere, L and van Steenberghe, D. One stage full- vs. partial-mouth disinfection in the treatment of patients with chronic adult or early-onset periodontitis. Part II: Long-term impact on microbial load. Journal of Periodontology 1999; 70:646-656. Quirynen, M., Mongardini, C., de Soete, M., et al. The role of chlorhexidine in the one-stage full-mouth disinfection treatment of patients with advanced adult periodontitis. Journal of Clinical Periodontology 2000; 27:578-589. Quirynen, M., De Soete, M., Boschmans, G., et al. “Benefit of onestage full-mouth disinfection” is explained by disinfection and root planing within 24 hours: a randomized controlled trial. Journal of Clinical Periodontology 2006; 33:639-647. Salvi, G.E. and Lang, N.P. Host response modulation in the management of periodontal diseases. Journal of Clinical Periodontology 2005; 32(Suppl. 6):108-129. Segelnick, S.L. and Weinberg, M.A. Reevaluation of initial therapy: When is the appropriate time? Journal of Periodontology 2006; 77:1598-1601. Serrano, C., Torres, N., Valdivieso, C., Castaño, C., Barrera, M. and Cabrales, M.A. Antibiotic resistance of periodontal pathogens obtained from frequent antibiotic users. Acta Odontologica Latinoamericana 2009; 22:99-104. Teles, R.P., Haffajee, A.D. and Socransky, S.S. Microbiological goals of periodontal therapy. Periodontology 2000, 2006; 42:180-208. Tunkel, J., Heinecke, A. and Flemming, T.F. A systematic review of efficacy of machine-driven and manual subgingival debridement in the treatment of chronic periodontitis. Journal of Clinical Periodontology 2002; 29(Suppl. 3):72-81. Van der Wejden, G.A. and Timmerman, M.F. A systematic review on the clinical efficacy of subgingival debridement in the treatment of chronic periodontitis. Journal of Clinical Periodontology 2002; 29(Suppl. 3):55-71. Van Winkelhoof, A.J., Van der Velden, U., Winkel, E.G. and de Graaf, J. Black-pigmented bacteroides and motile organisms on oral mucosal surfaces in individuals with and without periodontal breakdown. Journal of Periodontal Research 1986; 21:434-439. 22/12/2010 09:41:24 26 Journal of the International Academy of Periodontology (2011) 13/1 Van Winkelhoff, A.J., Tojhof, C.J. and de Graaf, J. Microbiological and clinical results of metronidazole plus amoxicillin therapy in Actinobacillus actinomycetemcomitans–associated periodontitis. Journal of Periodontology 1992; 63:52-57. Wennstrom, J.L., Tomasi, C., Bertele, A. and Dellasega, E. Full-mouth ultrasonic debridement versus quadrant scaling and root planing as an initial approach in the treatment of chronic periodontitis. Journal of Clinical Periodontology 2005; 32:851-859. Winkel, E.G., Van Winkelhoff, A.J., Timmerman, T.F., Van der Velden, U. and Van der Wejden, G.A. Amoxicillin plus metronidazole in the treatment of adult periodontitis patients. A double-blind placebo-controlled study. Journal of Clinical Periodontology 2001; 28:296-305. Zanatta, G.M., Bittencourt, S., Nociti Jr., F.H., Sallum, E.A., Sallum, A.W. and Casati, MZ. Periodontal debridement with povidoneiodine in periodontal treatment: Short-term clinical and biochemical observations. Journal of Periodontology 2006; 77:498-505. JIAP 10-003 Serrano.indd 26 22/12/2010 09:41:24 Journal of the International Academy of Periodontology 2011 13/1: 27–28 International Academy of Periodontology Lviv, Ukraine June 16-18, 2011 13th International Biennial Meeting Please complete form and return to: Congress Secretariat, 36 Pasichna Str, 79038 Lviv, Ukraine, Ms. Olena Uhryn phone: +38 (32) 2512039, fax: +38 (32) 2512151, e-mail: klinika@litech.lviv.ua Name: __________________________________________ Institution: __________________________________________ Mailing Address: __________________________________________ City: ___________________ Zip: _______________ Country: _____________ Telephone: ______________ Fax: ________________ e-mail:_______________ Participation in Gala-dinner YES NO Accompanying guest (Gala-dinner only) YES NO CONGRESS FEE-US dollars Dentists Dentists members of Ukrainian Oral Implantology Association and International Academy of Periodontology Dental technicians Dental technicians members of Ukrainian Oral Implantology association and International Academy of Periodontology Dental hygienists and dentists’ assistants Students BEFORE 02/15/2011 270.00 BEFORE 03/15/2011 300.00 AFTER 03/15/2011 350.00 250.00 280.00 300.00 170.00 200.00 250.00 150.00 180.00 230.00 120.00 50.00 150.00 75.00 170.00 100.00 DO YOU NEED OUR HELP IN HOTEL RESERVATION, PLEASE MARK YOUR CHOICE 1. 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UP TO 2X AS EFFECTIVE AS STRING FLOSS Improved Gingival Health Clinically proven up to twice as effective as string floss in reducing gingival bleeding Brushing & String after 14 days. Flossing Brushing & Waterpik® Water Flosser Independent clinical study. Data on file. NEW Plaque Seeker™ Tip Innovative new tip design to help remove even more stubborn plaque biofilm from hard to reach areas. For an abstract of the latest clinical research, please visit our website. www.waterpik.com Join the conversation at community.waterpik.com, and TM